DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
The instant application is a national stage entry under 35 U.S.C. § 371 of PCT/EP2021/059070 (filed 04/07/2021). Acknowledgement is made of Applicants’ claim for priority to foreign application EP20176939.5 (filed 05/27/2020).
Election/Restrictions
Applicant’s election without traverse of Group I in the reply filed on 11/14/2025 in response to a Restriction requirement is acknowledged.
Claims 52, 56, 60, 62, 64, 66, 70, and 72 have been cancelled by Applicants; claims 74-86, depending from claims 1 or 29 of Group I, have been added.
Claims 1-3, 7, 9, 20-21, 28-30, 34, 36, and 74-86 are now pending and examined on the merits herein.
Claim Interpretation
The following comments are made to establish broadest reasonable interpretation for the record.
The instant claims are drawn to viral vectors comprising myelo-specific and/or microglia-specific promoters (e.g., independent claims 1, 9) and fusion promoters comprising myelo-specific and microglia-specific promoters (e.g., independent claim 29). As evidenced by Greter, et al., microglia are a type of myeloid cell (pg. 1; par. 1). This is supported by the specification of the instant application, which discloses the promoters of the invention drive expression in myeloid cells and microglia (e.g., par. 2 of pg. 4, and par. 1 of pg. 5). Therefore, a promoter which reads on a microglia-specific promoter limitation of the instant claims necessarily reads as driving expression in both microglial cells and myeloid cells.
Claim Objections
Claims 30, 75, 81, and 85 are objected to because of the following informalities:
Claim 30 contains a typographical error: section 3) i) may be remedied by the following: “…SEQ ID NOs: 26-29[[:]]; or…”
Claim 75 appears to contain a typographical error, which may be remedied by the following: “The viral vector of claim 74, wherein the promoter…”
Claim 81 contains a typographical error, which may be remedied by the following: “The viral vector of claim 80, wherein the transcriptional activator…”
Claim 85 contains a typographical error in line 2, which may be remedied by the following: “…a VEGF-Receptor, an antagonist[[s]] to a metalloproteinase…”
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 2, 9, 20-21, 29-30, 74-77, and 86 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement.
The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The claims under rejection are directed to either viral vectors comprising myelo-specific and/or microglia-specific promoters (e.g., a miR223 promoter or a promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 1, or a functional fragment thereof, as recited in part a) of claim 2), as well as polypeptides encoded by a transgene of a viral vector of the invention (e.g., PGRN, or a functional fragment thereof; or a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9, or a functional fragment thereof, as recited in part (1) of claim 9), or to myelo-specific and/or microglial-specific promoters, per se (e.g. claim 29).
The issue at present is the scope imposed on the instant claims by the functional fragment limitation, as well as the 95% sequence identity limitation. This rejection will use the miR223 promoter limitation, i.e., SEQ ID NO: 1, for illustrative purposes.
Nucleic acid sequences for each promoter and transgene of the instant disclosure, and functional fragments thereof, are considered a genus of molecules. SEQ ID NO: 1 contains 778 nucleotides; thus, there are 2.63 X 1022 number of possible sequences with 95% sequence identity to SEQ ID NO: 1. As illustrated therein, the number of relevant species for just one promoter (i.e., just one of the genera of molecules) of the instant disclosure is extraordinarily large. This is further confounded by the ambiguity surrounding a clear definition of the functional fragment limitation of these claims; as recited in par. 3 of pg. 8 of the instant specification, “[a] functional fragment of the miR223 promoter is a nucleotide sequence comprising at least 20…consecutive nucleotides of SEQ ID NO: 1.” Thus, the issue at hand is whether or not the application provides sufficient written description for the genera of molecules covered by the claims.
To satisfy the written description aspect of 35 U.S.C. 112(a) for a claimed genus of molecules, it must be clear that: (1) the identifying characteristics of the claimed molecules have been disclosed, e.g., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed cor-relation between function and structure, or by a com-bination of such identifying characteristics; or (2) a representative number of species within the genus must be disclosed. See Eli Lilly, 119 F.3d at 1568, 43 USPQ2d at 1406.
Regarding (1): As discussed above, there are 2.63 X 1022 number of possible sequences in the genus of molecules comprising sequences with 95% sequence identity to SEQ ID NO: 1; Applicants do provide a physical description of the species of molecules thereof encoded by the nucleic acid sequence set forth in SEQ ID NO: 1. However, the issue is whether or not there is description of (i) the genus of molecules of sequences with 95% sequence identity to SEQ ID NO: 1, and (ii) the genus of molecules of functional fragments of SEQ ID NO: 1 or functional fragments of sequences sharing 95% sequence identity to SEQ ID NO: 1.
The instant disclosure provides limited guidance in this regard; e.g., “A promoter is said to have the functionality of the miR223 promoter, if it drives expression in the same cell types and at comparable levels and if it comprises at least a certain degree of sequence similarity to the miR223 promoter. A promoter is said to have a certain degree of sequence similarity to the miR223 promoter, if the promoter comprises a consecutive stretch of at least 20…nucleotides of SEQ ID NO: 1. Alternatively, a promoter is said to have a certain degree of sequence similarity to the miR223 promoter, if the promoter has at least 80%...sequence identity to the sequence shown in SEQ ID NO: 1.” (pgs. 8-9). However, this guidance lacks essential instruction; what substitutions, deletions, and/or insertions to SEQ ID NO: 1 may be made for a promoter to drive expression in the same cell types and at comparable levels? Do sequences identified in the prior art having at least 80% sequence identity to SEQ ID NO: 1 inherently drive expression in the same manner as the miR223 promoter of the instant invention? Would any 20 consecutive nucleotides of SEQ ID NO: 1 drive this expression, or is there some core structure within the sequence required for functionality? That is to say, which base pairs of SEQ ID NO: 1 are critical for the miR223 promoter to drive expression of a therapeutic protein in myeloid cells and microglia while being inactive in progenitor and/or stem cells? The disclosure lacks any clear guidance or teaching regarding which portions are required for the nucleic acid sequences of the instant invention; thus, there is no correlation of the function of these sequences and any particular structure.
Regarding (2): Applicants have failed to provide disclosure of species which are representative of the full scope of the claimed invention. Applicants’ disclosure is limited to the specific, fully defined sequences of the disclosure, as recited in the instant claims. That is, Applicants have disclosed a single species of the genus of molecules of sequences with 95% sequence identity to SEQ ID NO: 1; additionally, this single species is the sole species disclosed for the genus of molecules of functional fragments of SEQ ID NO: 1 and functional fragments of sequences sharing 95% sequence identity to SEQ ID NO: 1. This limited disclosure is not considered to be representative of the full breadth of the genera, as claimed.
Therefore, Applicants’ disclosure has support for an miR223 promoter comprising SEQ ID NO: 1, but not a promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 1, nor a functional fragment thereof. Likewise, Applicants’ have written description support for promoters and polypeptides encoded by their respective nucleic and amino acid sequences, but not promoters or polypeptides encoded by sequences having at least 95% sequence identity to their respective sequences, or functional fragments thereof.
Thus, a person having ordinary skill in the art would not be able to determine that Applicants were in possession of the invention, as claimed, at the time the invention was made. Accordingly, the claims are considered to lack sufficient written description and are properly rejected under 35 U.S.C. § 112(a).
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 21 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claim 21: In part 4) of claim 21, the limitations recited in (i)-(v) do not clearly or precisely define the metes and bounds of the invention; specifically, the SEQ ID NOs do not appear to match the associated promoter. For example, (i) recites, “…a TMEM119 promoter, or a promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 7…”. However, according to the Sequence listing, SEQ ID NO: 5 is an AIF1 promoter sequence, SEQ ID NO: 6 is an ITGAM promoter sequence, and SEQ ID NO: 7 is a Progranulin isoform 1 sequence.
Likewise, while SEQ ID NO: 2 is a P2RY12 promoter sequence, the remaining sequences recited in (ii) are for a TMEM119 promoter sequence (SEQ ID NO: 3) and an OLFML3 promoter sequence (SEQ ID NO: 4); (iii) recites an OLFML3 promoter, but lists SEQ ID NOs: 8 and 9, which encode Progranulin isoforms 2 and 3, respectively; (iv) recites an ITGAM promoter, but SEQ ID NO: 11 encodes human IL-12 subunit alpha; and (v) recites an AIF1 promoter, but SEQ ID NO: 10 encodes human interferon-gamma.
The metes and bounds of the instant claim are not clearly or precisely defined because, as currently written, it would not be understood by a person having ordinary skill in the art what the miR223 promoter of part 4) should be operably linked to. For instance, are the sequences recited in the limitation of (i) typographical errors (i.e., copied and pasted in error)? Or does (i) recite an improper Markush group, reciting a TMEM119 promoter, an AIF1 promoter sequence, an ITGAM promoter sequence, and a Progranulin isoform 1 sequence as alternative members?
As currently written, the lack of clear and defined metes and bounds render the instant claim indefinite; thus, claim 21 is rejected under 35 U.S.C. § 112(b).
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1-3, 7, 28, 34, 36, and 78 are rejected under 35 U.S.C. 102(a)(2) as being anticipated by Genkin, et al. (US 2022/0241394).
Genkin, et al. (hereinafter Genkin) teaches nervous system-specific delivery and/or expression of an enzyme which has a deoxyribonuclease activity in cerebrospinal fluid, brain, and other parts of the nervous system (Abstract).
Regarding claims 1-2: Genkin teaches a recombinant adeno-associated virus (rAAV) expression vector comprising a capsid protein and nucleic acid comprising a nervous system-specific promoter operably linked to a nucleotide sequence encoding an enzyme with deoxyribonuclease (DNase) activity (par. 0040). Genkin teaches expression of an enzyme having DNase activity for enhanced clearance of microbial and viral cell free DNA accumulated in cerebrospinal fluid, brain, and other parts of the nervous system is useful for the treatment of various diseases associated with protein misfolding (par. 0003). This reads on the viral vector comprising a nucleic acid molecule encoding a therapeutic polypeptide or a combination of therapeutic polypeptides under control of a promoter or promoter fragment limitation recited in claim 1.
Further disclosed is an embodiment wherein the nervous system-specific promoter is a TMEM119 microglia-specific promoter (par. 0041), comprising SEQ ID NO: 39 (par. 0015). SEQ ID NO: 39 shares 100% sequence identity with instant SEQ ID NO: 3, 99.6% sequence identity with instant SEQ ID NO: 23, and 99.4% sequence identity with instant SEQ ID NO: 24; please see end of Office action for sequence alignments. Thus, this reads on the remaining wherein the promoter or promoter fragment drives expression of the therapeutic protein or the combination of therapeutic proteins in myeloid cells and microglia, and wherein the promoter or promoter fragment is inactive in progenitor and/or stem cells limitation recited in claim 1, as well as the wherein the promoter is a TMEM119 promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 3, SEQ ID NO: 23, or SEQ ID NO: 24 limitation recited in claim 2.
Regarding claims 3, 7: Genkin teaches an embodiment wherein the enzyme having DNase activity is Granzyme B (par. 0017); this reads on the wherein the therapeutic polypeptide is a polypeptide that restores a cellular function and/or elicits a cellular response in a cell limitation recited in claim 3, as well as the wherein the polypeptide that restores a cellular function and/or elicits a cellular response in a cell comprises Granzyme B limitation recited in claim 7.
Regarding claim 28: The rAAV vector of Genkin (par. 0040) reads on the wherein the viral vector is an adeno- associated viral vector limitation recited in claim 28.
Regarding claim 34: Genkin teaches an embodiment wherein a packaging cell comprises the rAAV vector for production of viral particles (par. 0196); the packaging cell reads on the host cell comprising the viral vector limitation recited in claim 34.
Regarding claim 36: Genkin teaches an embodiment wherein the rAAV vector is formulated in a pharmaceutical composition (par. 0271); this reads on the pharmaceutical composition comprising the viral vector limitation recited in claim 36.
Regarding claim 78: Genkin further discloses the rAAV may further comprise a woodchuck hepatitis post-transcriptional regulatory element, or WPRE (par. 0023); this reads on the wherein the viral vector comprises at least one transcriptional regulatory element limitation recited in claim 78.
Claims 9 and 20-21 are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by Chen Plotkin, et al. (WO 2017/151884).
Chen Plotkin, et al. (hereinafter Chen Plotkin) teaches methods and uses for delivering progranulin to the central nervous system of a mammal (Abstract).
Regarding claim 9: Chen Plotkin teaches an rAAV vector comprising a nucleic acid encoding progranulin (par. 0009), wherein the vector comprises a promoter (par. 0015). The disclosure also teaches an embodiment wherein progranulin is encoded by SEQ ID NO: 1 (par. 0105), which shares 100% sequence identity with instant SEQ ID NO: 7 and 95% sequence identity with SEQ ID NO: 8; please see end of Office action for sequence alignments. Therefore, this reads on the viral vector comprising a transgene under control of one or more promoters, wherein the transgene encodes PGRN or a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 7 or SEQ ID NO: 8 limitation recited in claim 9.
Regarding claim 20: Following the above discussion, the rAAV of Chen Plotkin reads on the wherein the viral vector is an adeno-associated viral vector limitation recited in claim 20. Chen Plotkin teaches an embodiment wherein the vector further comprises one or more expression control regulatory elements, e.g., transcription initiation sequences (par. 0152); this reads on the wherein the viral vector comprises at least one transcriptional regulatory element, and wherein said at least one transcriptional regulatory element is arranged such that it activates a transcriptional activity of the promoter limitation recited in claim 20.
Regarding claim 21: Following the above discussion, Chen Plotkin teaches the regulatory element may comprise DNA sequences involved in the binding of protein factors that modulate/control effectiveness of transcription initiation (par. 0155); this reads on the wherein the at least one transcriptional regulatory element comprises a binding site for a transcriptional activator or repressor limitation recited in claim 21.
Claims 9 and 20-21 are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by Beech, et al. (US 2018/0333486).
Beech, et al. (hereinafter Beech) teaches methods of generating in vitro engineered immune cells conditionally expressing IL-12 and one or more immunomodulators (Abstract).
Regarding claims 9, 20: Beech teaches a vector comprising a polynucleotide encoding one or more immunomodulators and a polynucleotide encoding IL-12, linked to a promoter activated by a ligand-dependent transcription factor (pars. 0010, 0461-0462), wherein the one or more immunomodulators are IL-2, IL-15, IL-21, GM-CSF, IFN-alpha, IFN-gamma (par. 0009). Beech additionally teaches the vector as a recombinant adenovirus vector (par. 0008).
Beech further discloses the following embodiments:
wherein IL-12 is encoded by SEQ ID NO: 24 (par. 0191), which shares 98.9% sequence identity to instant SEQ ID NO: 11,
wherein IL-2 is encoded by the sequence set forth in Accession No. AAA70092 (par. 0073), which shares 96% sequence identity to instant SEQ ID NO: 17,
wherein IL-15 is encoded by SEQ ID NO: 20 (par. 0187), which shares 100% sequence identity to instant SEQ ID NO: 18,
wherein IL-21 is encoded by SEQ ID NO: 18 (par. 0185), which shares 100% sequence identity to instant SEQ ID NO: 19,
wherein GM-CSF is encoded by the sequence set forth in Accession No. AAA52122 (par. 0124), which shares 100% sequence identity to instant SEQ ID NO: 13, and
wherein IFN-gamma is encoded by the sequence set forth in Accession No. NP_000610 (par. 0096), which shares 100% sequence identity to instant SEQ ID NO: 10. Please see end of Office action for sequence alignments.
These read on the wherein the transgene encodes a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 11, a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 10, a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 13, a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 17, a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 18, a polypeptide having at least 95% sequence identity to the sequence of SEQ ID NO: 19 limitations recited in parts (2)(b), (3)(b), (4)(b), (8)(b), (9)(b), (10)(b), and (11)(b) of claim 9.
Thus, Beech anticipates the viral vector comprising a transgene under control of one or more promoters, wherein the transgene encodes IL-12, IFN-gamma, GM-CSF, GM-CSF and IFN-gamma, IL-2, IL-15, IL-21, and IFN-alpha limitations recited in parts (2)(a), (3)(a), (4)(a), (6)(a), (8)(a), (9)(a), (10)(a), and (11)(a) of claim 9. Additionally, this anticipates the wherein the viral vector comprises at least one transcriptional regulatory element, and wherein said at least one transcriptional regulatory element is arranged such that it activates a transcriptional activity of the promoter and the wherein the viral vector is an adenoviral vector limitations recited in claim 20.
Regarding claim 21: Following the above discussion, the ligand-dependent transcription factor which activates the promoter taught by Beech (par. 0010) anticipates the wherein the at least one transcriptional regulatory element comprises a binding site for a transcriptional activator limitation recited in claim 21.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-3, 7, 9, 20-21, 28, 34, 36, and 78-80 are rejected under 35 U.S.C. 103 as being unpatentable over Chen Plotkin, et al. (WO 2017/151884), in view of Genkin, et al. (US 2022/0241394).
The teachings of Chen Plotkin and Genkin are set forth above. Claims 9 and 20-21 are anticipated by Chen Plotkin.
Regarding claims 1-2: Chen Plotkin teaches an rAAV vector comprising a nucleic acid encoding progranulin (par. 0009), wherein the vector comprises a promoter; e.g., a chicken beta actin promoter or CAG promoter (par. 0015). Chen Plotkin does not teach the promoter as a myeloid-restricted or microglia-specific promoter.
However, Genkin teaches a TMEM119 microglia-specific promoter (par. 0041), comprising SEQ ID NO: 39 (par. 0015). SEQ ID NO: 39 shares 100% sequence identity with instant SEQ ID NO: 3, 99.6% sequence identity with instant SEQ ID NO: 23, and 99.4% sequence identity with instant SEQ ID NO: 24; please see end of Office action for sequence alignments. This reads on the wherein the promoter is a TMEM119 promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 3, SEQ ID NO: 23 or SEQ ID NO: 24 limitation recited in claim 2.
It would have been prima facie obvious to a person having ordinary skill in the art to have modified the rAAV of Chen Plotkin by substituting the chicken beta actin or CAG promoter with the microglia-specific TMEM119 promoter as taught by Genkin. This conclusion of obviousness is based on the ‘substitution rationale’; the use of the microglia-specific TMEM119 promoter in place of the chicken beta actin or CAG promoter is a predictable use of prior art elements according to their established functions of driving transgene expression in cells, leading to the predictable result of PGRN expression in microglia. Chen Plotkin teaches an embodiment wherein the rAAV is used to transduce CNS cells such as glial cells (par. 0021), which, as is well-known in the art, include microglia; thus, one skilled in the art would have more than a reasonable expectation of success in such a modification. This rationale aligns with the principle of a simple substitution of one known element for another to obtain predictable results; see MPEP 2143(I)(B).
Thus, the modified rAAV of Chen Plotkin reads on the viral vector comprising a nucleic acid molecule encoding a therapeutic polypeptide or a combination of therapeutic polypeptides under control of a promoter or promoter fragment, wherein the promoter or promoter fragment drives expression of the therapeutic protein or the combination of therapeutic proteins in myeloid cells and microglia, and wherein the promoter or promoter fragment is inactive in progenitor and/or stem cells limitation recited in claim 1.
Regarding claims 3, 7: Following the above discussion, the modified rAAV of Chen Plotkin encoding progranulin (par. 0009) reads on the wherein the therapeutic polypeptide is a polypeptide that restores a cellular function and/or elicits a cellular response in a cell limitation recited in claim 3, as well as the wherein the polypeptide that restores a cellular function and/or elicits a cellular response in a cell comprises progranulin limitation recited in claim 7.
Regarding claim 28: Following the above discussion, the modified rAAV of Chen Plotkin reads on the wherein the viral vector is an adeno-associated viral vector limitation recited in claim 28.
Regarding claim 34: Following the above discussion, Chen Plotkin teaches an embodiment wherein transduction-competent AAV particles are produced in a host cell (par. 0101); this reads on the host cell comprising the viral vector according to claim 1 limitation recited in claim 34.
Regarding claim 36: Following the above discussion, Chen Plotkin teaches the rAAV can be formulated in a pharmaceutically acceptable formulation for administration (par. 0116); this reads on the pharmaceutical composition comprising the viral vector according to claim 1 limitation recited in claim 36.
Regarding claims 78-80: Following the above discussion, Chen Plotkin teaches an embodiment wherein the vector further comprises one or more expression control regulatory elements, e.g., transcription initiation sequences (par. 0152), wherein the regulatory element may comprise DNA sequences involved in the binding of protein factors that modulate/control effectiveness of transcription initiation (par. 0155); this reads on the wherein the viral vector comprises at least one transcriptional regulatory element limitation recited in claim 78, the wherein the at least one transcriptional regulatory element is arranged such that it inhibits or activates a transcriptional activity of the promoter limitation recited in claim 79, and the wherein the at least one transcriptional regulatory element comprises a binding site for a transcriptional activator or repressor limitation recited in claim 80.
Claims 9, 20-21, 29, 74-77, and 86 are rejected under 35 U.S.C. 103 as being unpatentable over Chen Plotkin, et al. (WO 2017/151884), in view of Genkin, et al. (US 2022/0241394) and Brendel, et al. (Hum Gene Ther Methods. 2013), further in view of Yoshihito, et al. (Neurosurgery. 2007); and as evidenced by Herz, et al. (Immunity. 2017), and Harding, et al. (EMBO J. 1989).
The teachings of Chen Plotkin and Genkin are set forth above. Claims 9 and 20-21 are anticipated by Chen Plotkin.
Brendel, et al. (hereinafter Brendel) teaches myelospecificity of the human miR223 promoter (Abstract). Yoshihito, et al. (hereinafter Yoshihito) teaches the intracerebral transplantation of VEGF gene-transferred bone marrow stromal cells (Abstract).
Regarding claims 29, 74-77, 86: The modified rAAV vector of Chen Plotkin comprising the TMEM119 microglia-specific promoter of Genkin is set forth above. Following on from that discussion, the modified rAAV vector does not comprise a fusion promoter comprising a myelo-specific miR223 promoter operably linked to the microglia-specific promoter.
However, Chen Plotkin teaches an embodiment wherein the rAAV is used to transduce central nervous system (CNS) cells including glial cells and meningeal cells (par. 0021); it is well-known in the art microglia are glial cells. As evidenced by Herz, et al., meningeal cells include a diverse population of myeloid cells, such as meningeal macrophages (pg. 945; col. 2, par. 2), monocytes (pg. 946; col. 1, par. 3), dendritic cells (pg. 946; col. 1, par. 5), and granulocytes (pg. 946; col. 2, par. 2).
Brendel teaches a lentiviral construct comprising an miR223 promoter and p47phox or gp91phox transgene (pg. 152; par. 4). Brendel teaches the miR223 promoter for myeloid-restricted transgene expression (pg. 156; col. 2, par. 1); e.g., the human myelomonocytic X-CGD cell line (Fig. 1; Fig. 2A, upper quartiles), macrophages (Fig. 4), granulocytes (Fig. 5A). Brendel further teaches the miR223 promoter has only marginal activity in hematopoietic stem and progenitor cells (Abstract; pg. 155, par. 1; Fig. 2A, lower quartiles; Fig. 3).
It would have been prima facie obvious to a person having ordinary skill in the art to have further modified the rAAV of Chen Plotkin by incorporating a fusion promoter wherein the myelo-specific miR223 promoter of Brendel is operably linked to the TMEM119 microglia-specific promoter. This conclusion of obviousness is based on the ‘combining known alternatives rationale’. As Chen Plotkin teaches the rAAV encoding progranulin as appropriate for use in glial and meningeal cells of the CNS (e.g., microglia and myeloid cells, respectively), one skilled in the art would be motivated to incorporate a fusion promoter comprising the miR223 promoter and TMEM119 promoter with the expectation of increasing transduction efficiency due to an additive effect of the two promoters. The combination of the myelo-specific and microglia-specific promoters is a predictable use of known alternatives for driving transgene expression in CNS cells, leading to the predictable result of PGRN expression in microglia and myeloid cells of the CNS. See MPEP 2143(I)(A). As evidenced by Harding, et al. (“Results”; pgs. 1206-1207), techniques for the generation of fusion promoters are well-known in the art. Chen Plotkin also discloses the use of hybrid promoters (par. 0156). Additionally, Yoshihito discloses a viral vector comprising a gene of interest of the control of fusion promoters for intracerebral transplantation (pg. 587; pars. 3, 6). Thus, a person having ordinary skill in the art would have more than a reasonable expectation of success in the generation of a fusion promoter for delivery of progranulin to microglia and myeloid cells of the CNS.
Therefore, the modified rAAV of Chen Plotkin as set forth above renders obvious the fusion promoter comprising a miR223 promoter and a microglia-specific promoter, wherein the miR223 promoter is operably linked to the microglia-specific promoter limitation recited in claim 29, the wherein the promoter is a fusion promoter comprising a miR223 promoter limitation recited in claim 74, and the wherein the promoter is operably linked to a microglia-specific promoter limitation recited in claim 75.
Additionally, as set forth above, Genkin teaches the TMEM119 promoter comprises SEQ ID NO: 39 (par. 0015), which shares 100% sequence identity with instant SEQ ID NO: 3, 99.6% sequence identity with instant SEQ ID NO: 23, and 99.4% sequence identity with instant SEQ ID NO: 24; please see end of Office action for sequence alignments. Thus, the modified lentiviral construct of Chen Plotkin as set forth above renders obvious the wherein the microglia-specific promoter is a TMEM119 promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 3, SEQ ID NO: 23, or SEQ ID NO: 24 limitation recited in claim 76, the wherein the promoter is a fusion promoter comprising a miR223 promoter operably linked to a TMEM119 promoter, or a promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 3, SEQ ID NO: 23 or SEQ ID NO: 24 limitation recited in claim 77, and the fusion promoter comprising a miR223 promoter and a microglia-specific promoter, wherein the microglia-specific promoter is a TMEM119 promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 3, SEQ ID NO: 23, or SEQ ID NO: 24, wherein the miR223 promoter is operably linked to the microglia-specific promoter limitation recited in claim 86.
Claims 1-3, 7, 28, 34, 36, and 78-81 are rejected under 35 U.S.C. 103 as being unpatentable over Genkin, et al. (US 2022/0241394), in view of Wilson, et al. (US 2017/0000904).
The teachings of Genkin are set forth above. Claims 1-3, 7, 28, 34, 36, 78 are anticipated by Genkin.
Wilson, et al. (hereinafter Wilson) teaches pharmacologically induced transgene ablation, wherein replication-deficient viruses are used to deliver a transgene encoding a therapeutic product so that it is expressed in the subject, but can be reversibly or irreversibly turned off by administering a pharmacological agent (Abstract).
Regarding claims 79-81: It is set forth above Genkin anticipates the viral vector of claim 78. Genkin does not teach the limitations recited in the instant claims.
Wilson teaches a pharmacologically induced transgene ablation (PITA) system designed for the delivery of a transgene, wherein a therapeutic gene product may be permanently or temporarily ablated in response to a pharmacological agent (par. 0062). In one embodiment, the PITA system comprises a transgene unit encoding a therapeutic antibody, an ablation unit encoding an endonuclease that is in operative association with a transcription factor inducible promoter, and a dimerizable transcription factor domain unit (par. 0045). Prior to administration of rapamycin or a rapalog, there is baseline expression of the therapeutic antibody and of the two transcription factor domain fusion proteins; upon rapamycin administration, the dimerized transcription factor induces expression of the endonuclease, which cleaves the endonuclease recognition domain in the transgene unit, thereby ablating transgene expression (par. 0045; Fig. 17). This reads on the wherein the at least one transcriptional regulatory element is arranged such that it inhibits or activates a transcriptional activity of the promoter limitation recited in claim 79, the wherein the at least one transcriptional regulatory element comprises a binding site for a transcriptional activator or repressor limitation recited in claim 80, and the wherein the transcriptional activator or repressor comprises a dimerizer system limitation recited in claim 81.
It would have been prima facie obvious to a person having ordinary skill in the art to have modified the viral vector of Genkin by including the PITA system as taught by Wilson. This conclusion of obviousness is based on the ‘teaching, suggestion, or motivation rationale’; one would be motivated to do so because, as taught by Wilson, there is a need to be able to turn off or permanently ablate transgene expression if it is no longer needed or if toxicity due to long-term drug administration ensues (par. 0004). Further, as Wilson teaches the PITA system for use in AAV-mediated delivery of a therapeutic product (par. 0008), one skilled in the art would have more than a reasonable expectation of success in incorporating the PITA system into the rAAV of Genkin.
Thus, the modified viral vector of Genkin renders obvious the limitations recited in claims 79-81.
Claims 1-3, 7, 28, 34, 36, 78, and 82 are rejected under 35 U.S.C. 103 as being unpatentable over Genkin, et al. (US 2022/0241394), in view of Strobel, et al. (ACS Synth Biol. 2020).
The teachings of Genkin are set forth above. Claims 1-3, 7, 28, 34, 36, 78 are anticipated by Genkin.
Strobel, et al. (hereinafter Strobel) teaches the utilization of the tetracycline-dependent ribozyme K19 for controlling AAV-mediated transgene expression (Abstract).
Regarding claim 82: It is set forth above Genkin anticipates the viral vector of claim 1. Genkin does not teach the vector as encoding a riboswitch, as required by the limitation recited in the instant claim.
Strobel teaches inclusion of the tetracycline-dependent ribozyme K19 riboswitch in an AAV vector repressed transgene expression to 3.1% of constitutive control levels, whereas maximal, 13.2-fold induction reached 40.1% 8 hours after tetracycline administration (pg. 1299; par. 2; Fig. 6). Upon a single administration of increasing tetracycline doses (3, 10, 30, 90 mg/kg), transgene expression was rapidly induced to dose-dependent peak expression levels of approximately 12, 16, 28, and 30% of control, respectively; the duration of transgene induction was dose-dependent, with a return to baseline occurring more rapidly at lower doses (pg. 1299; pars. 4-5; Fig. 7b). Further, after waiting one week to ensure complete tetracycline clearance and to stimulate a treatment-free phase with desired expression shutdown, transgene expression had returned to baseline; however, upon re-administration, expression was induced in the same dose-dependent fashion as seen previously, reaching a maximum of 34% of control levels (i.e., 14.7-fold induction) at the highest ligand dose (pg. 1299; pars. 4-5; Fig. 7b).
It would have been prima facie obvious to a person having ordinary skill in the art to have included the tetracycline-dependent ribozyme K19 riboswitch of Strobel in the rAAV construct of Genkin. This conclusion of obviousness is based on the ‘teaching, suggestion, or motivation rationale’. One would be motivated to do so because, as disclosed by Strobel, the possibility to dose-dependently adjust expression levels, potentially enabling tuning of therapeutic effects in a patient-centric fashion, is particularly important given the fluctuations in expression often observed across patients in gene therapy trials, as well as detrimental effects caused by the uncontrolled expression of some therapeutic proteins (pg. 1300; par. 3). Further, as Strobel teaches the use of the riboswitch in an AAV vector (pg. 1299; par. 2), one skilled in the art would have a reasonable expectation of success in its use in the rAAV of Genkin.
This renders obvious the limitations recited in claim 82.
Claims 1-3, 7, 9, 20-21, 28, 34, 36, 78-80, and 83-85 are rejected under 35 U.S.C. 103 as being unpatentable over Beech, et al. (US 2018/0333486) in view of Genkin, et al. (US 2022/0241394).
The teachings of Beech and Genkin are set forth above. Claims 9 and 20-21 are anticipated by Beech.
Regarding claims 1-2: It is set forth above Beech teaches a recombinant adenovirus vector comprising a polynucleotide encoding IL-12 and one or more of IL-2, IL-15, IL-21, GM-CSF, IFN-alpha, IFN-gamma, linked to a promoter activated by a ligand-dependent transcription factor (pars. 0008-0010, 0462). Beech does not teach the promoter limitation of the instant claims; however, disclosed is an embodiment wherein the promoter is the beta-globin gene control region active in myeloid cells (par. 0233).
Genkin teaches a TMEM119 microglia-specific promoter (par. 0041), comprising SEQ ID NO: 39 (par. 0015); SEQ ID NO: 39 shares 100% sequence identity with instant SEQ ID NO: 3, 99.6% sequence identity with instant SEQ ID NO: 23, and 99.4% sequence identity with instant SEQ ID NO: 24. Please see end of Office action for sequence alignments.
It would have been prima facie obvious to a person having ordinary skill in the art to have modified the vector of Beech by substituting the beta-globin gene control region promoter with the TMEM119 promoter taught by Genkin. This conclusion of obviousness is based on the ‘substitution rationale’. The use of the TMEM119 promoter in place of the beta-globin gene control region promoter is a predictable use of prior art elements according to their established functions of driving transgene expression in a cell, leading to the predictable result of expression of IL-12 and one or more of IL-2, IL-15, IL-21, GM-CSF, IFN-alpha, IFN-gamma. As Beech teaches the use of a promoter active in myeloid cells, and the TMEM119 promoter of Genkin drives expression in microglia (a myeloid cell), a person skilled in the art would have more than a reasonable expectation of success. This rationale aligns with the principle of a simple substitution of one known element for another to obtain predictable results; see MPEP 2143(I)(B).
Therefore, the modified vector of Beech as set forth above renders obvious the viral vector comprising a nucleic acid molecule encoding a therapeutic polypeptide or a combination of therapeutic polypeptides under control of a promoter or promoter fragment, wherein the promoter or promoter fragment drives expression of the therapeutic protein or the combination of therapeutic proteins in myeloid cells and microglia, and wherein the promoter or promoter fragment is inactive in progenitor and/or stem cells limitations recited in claim 1, as well as the wherein the promoter is a miR223 promoter limitation recited in claim 2.
Regarding claims 3, 7: Following the above discussion, the modified vector of Beech renders obvious the wherein the therapeutic polypeptide is a polypeptide that restores a cellular function and/or elicits a cellular response in a cell limitation recited in claim 3, as well as the wherein the polypeptide that restores a cellular function and/or elicits a cellular response in a cell comprises IL-2, IL-12, IL-15, IL-21, IFN-alpha, IFN-gamma, and GM-CSF limitations recited in claim 7.
Regarding claim 28: Following the above discussion, the modified adenoviral vector of Beech reads on the wherein the viral vector is an adenoviral vector limitation recited in claim 28.
Regarding claims 34, 36: Following the above discussion, Beech teaches engineered cells which conditionally express the IL-12 and immunomodulator proteins, wherein the engineered immune cells are formulated into a pharmaceutical composition (par. 0456); this reads on the host cell comprising the viral vector according to claim 1 limitation recited in claim 34, as well as the pharmaceutical composition comprising the viral vector according to claim 1 and/or a host cell comprising the viral vector according to claim 1 limitations recited in claim 36.
Regarding claims 78-80: Following the above discussion, the ligand-dependent transcription factor which activates the promoter (par. 0010) disclosed by Beech reads on the wherein the viral vector comprises at least one transcriptional regulatory element limitation recited in claim 78, the wherein the at least one transcriptional regulatory element is arranged such that it activates a transcriptional activity of the promoter limitation recited in claim 79, as well as the wherein the at least one transcriptional regulatory element comprises a binding site for a transcriptional activator limitation recited in claim 80.
Regarding claim 83: Following the above discussion, Beech discloses dendritic cells (DC) engineered to produce IL-12p70 via recombinant adenovirus infection dramatically improved cross-priming of a broadly-reactive, tumor-specific CD8+ T cell repertoire in concert with tumor rejection (par. 0007). Thus, the modified vector of Beech encoding IL-12 reads on the wherein the therapeutic polypeptide is a polypeptide that enables and/or increases target specificity of a cell limitation recited in claim 83.
Regarding claim 84-85: Following the above discussion, Beech teaches a method of producing a population of immune cells or therapy support cells (TSC) by modifying the cells with the recombinant vector (par. 0011); prior to intratumoral injection, these cells may be treated with a factor to stimulate their activity, such as CD40 or CD40L (par. 0486). This renders obvious the wherein the polypeptide that enables and/or increases target specificity of a cell enables and/or increases specificity to a tumor antigen limitation recited in claim 84, as well as the wherein the tumor antigen is CD40/CD40L limitation recited in claim 85.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
Claims 1-3, 28, and 34 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1 of copending Application No. 17/765,172 (reference application).
Although the claims at issue are not identical, they are not patentably distinct from each other.
Regarding claims 1-3, 28, 34: Claim 1 of copending Application No. 17/765,172 recites a lentiviral vector comprising a coding nucleic acid sequence encoding a polypeptide selected from gp91phox and p47phox; under transcriptional control of a miR223 promoter sequence comprising the sequence of SEQ ID NO: 1. As copending SEQ ID NO: 1 shares 100% sequence identity with instant SEQ ID NO: 1, this reads on the viral vector comprising a nucleic acid molecule encoding a therapeutic polypeptide or a combination of therapeutic polypeptides under control of a promoter or promoter fragment, wherein the promoter or promoter fragment drives expression of the therapeutic protein or the combination of therapeutic proteins in myeloid cells and microglia, and wherein the promoter or promoter fragment is inactive in progenitor and/or stem cells limitation recited in instant claim 1, the wherein the promoter is a miR223 promoter having at least 95% sequence identity to the sequence of SEQ ID NO: 1 limitation recited in instant claim 2, the wherein the therapeutic polypeptide is a polypeptide that restores a cellular function and/or elicits a cellular response in a cell limitation recited in instant claim 3, the wherein the viral vector is a retroviral vector limitation recited in instant claim 28, and the host cell comprising the viral vector according to claim 1 limitation recited in instant claim 34.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Allowable Subject Matter
All claims are rejected. However, in the interest of advancing prosecution, the following is a statement of reasons for the indication of allowable subject matter:
During the course of examination, the fusion promoter sequences of the instant application as set forth in SEQ ID NOs: 26-29 were found to be non-obvious in view of the prior art.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to GINA PRONZATI whose telephone number is (571)270-5725. The examiner can normally be reached Monday - Friday 9:00a - 5:00p ET.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, CHRISTOPHER BABIC can be reached at (571)272-8507. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/GINA PRONZATI/Examiner, Art Unit 1633
/ALLISON M FOX/Primary Examiner, Art Unit 1633
SEQUENCE ALIGNMENTS FOR GENKIN (US 2022/0241394)
Query, ‘Qy’ (SEQ ID NO: 3) vs. Database, ‘Db’ (Genkin; SEQ ID NO: 39)
PNG
media_image1.png
272
653
media_image1.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 23) vs. Database, ‘Db’ (Genkin; SEQ ID NO: 39)
PNG
media_image2.png
275
656
media_image2.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 24) vs. Database, ‘Db’ (Genkin; SEQ ID NO: 39)
PNG
media_image3.png
279
650
media_image3.png
Greyscale
Continued on next page.
SEQUENCE ALIGNMENTS FOR CHEN PLOTKIN (WO 2017/151884)
Query, ‘Qy’ (SEQ ID NO: 7) vs. Database, ‘Db’ (Chen Plotkin; SEQ ID NO: 1)
PNG
media_image4.png
296
589
media_image4.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 8) vs. Database, ‘Db’ (Chen Plotkin; SEQ ID NO: 1)
PNG
media_image5.png
293
601
media_image5.png
Greyscale
Continued on next page.
SEQUENCE ALIGNMENTS FOR BEECH (US 2018/0333486)
Query, ‘Qy’ (SEQ ID NO: 18) vs. Database, ‘Db’ (Beech; SEQ ID NO: 20)
PNG
media_image6.png
273
641
media_image6.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 19) vs. Database, ‘Db’ (Beech; SEQ ID NO: 18)
PNG
media_image7.png
277
649
media_image7.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 11) vs. Database, ‘Db’ (Beech; SEQ ID NO: 24)
PNG
media_image8.png
335
651
media_image8.png
Greyscale
Continued on next page.
SEQUENCE ALIGNMENTS FOR BEECH (US 2018/0333486) (continued)
Query, ‘Qy’ (SEQ ID NO: 17) vs. Database, ‘Db’ (Beech; Accession No. AAA70092)
PNG
media_image9.png
297
613
media_image9.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 13) vs. Database, ‘Db’ (Beech; Accession No. AAA52122)
PNG
media_image10.png
327
628
media_image10.png
Greyscale
Query, ‘Qy’ (SEQ ID NO: 10) vs. Database, ‘Db’ (Beech; Accession No. NP_000610)
PNG
media_image11.png
329
626
media_image11.png
Greyscale
Continued on next page.
SEQUENCE ALIGNMENTS FOR APPLICATION NO. 17/765,172
Query, ‘Qy’ (SEQ ID NO: 1) vs. Database, ‘Db’ (copending Application No. 17/765,172; SEQ ID NO: 1)
PNG
media_image12.png
272
648
media_image12.png
Greyscale