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
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 4/14/2023 and 11/7/2025 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
Election/Restrictions
Applicant’s election without traverse of Group I drawn to methods for deaminating an adenosine (A) in an USH2A gene in the reply filed on 11/7/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
Status of claims
Claims 1-2, 6-7, 10, 14, 16, 18, 22, 23, 26, 28, 32-37, 76 and 114 are pending and under examination. Claims 38, 45, 90, 95-96, 103, and 108 are pending and withdrawn from consideration. Claims 3-5, 8-9, 11-13, 15, 17, 19-21, 24-25, 27, 29-31, 39-44, 46-75, 77-89, 91-94, 97-102, 104-107, and 109-113 are cancelled.
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.
Claims 1-2, 6-7, 10, 14, 16, 18 , 22, 23 , 26, 28, 32-37, and 76 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as failing to set forth 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.
Claim 1 requires a target nucleic acid sequence in the USH2A gene having at least 90% identity to any one of SEQ ID NOs: 12-35 or naturally occurring variants. It is not clear from the claims or the specification regarding what naturally occurring variants are envisioned or encompassed. Further the structure or sequence of the variants are unclear to a person of ordinary skill in the art. Appropriate clarification is required.
Claim 7 required an adenosine deaminase comprising amino acid of SEQ ID NO: 79, 90 and 91. However the specification recites that “Streptococcus pyogenes Cas9 as provided by any one of SEQ ID NOs: 74, 77, and 79.” See Specification [0477]. Also see Table in page 66 for SEQ ID NOs 90-91. It is not clear if SEQ ID NO: 79, 90 and 91 are Adenosine deaminase or Cas9, which are entirely different proteins. Appropriate clarification is required. For the sake of compact prosecution, it is assumed that the claim requires a Cas protein having an amino acid sequence having at least 90% to any one of SEQ ID NOs: 79, 90, and 91.
Claims 2, 6-7, 10, 14, 16, 18 , 22, 23 , 26, 28, 32-37, 76 and 114 are rejected for their dependency on rejected claims.
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 1-2, 6-7, 10, 14, 16, 18 , 22, 23 , 26, 28, 32-37, and 76 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.
Regarding claim 1: Applicant’s claims do not provide support for all nucleotides comprising 90% identity to SEQ ID NO: 12-35. The specification as filed does not provide sufficient evidence that Applicants were in possession of the full scope of the claimed invention at the time of filing of the instant invention. As such, >90% identity to a 51 base pair polynucleotide, polynucleotide encompasses
51
C
5
×
3
5
= more than
5.7
×
10
8
nucleotide molecules. This amounts to enormous number of polynucleotides which do not have support in the specification. The specification also does not provide any guidance on how or where the nucleotides need to be changed. Thus, Applicants were not in possession of the full scope of the claimed invention at the time of filing of the instant invention. It is clear that the vector of the invention was restricted to SEQ ID NOs: 12-35. Similar rejection applies to claims 18 and 26.
Regarding claims 2 and 7: Applicant’s claims do not provide support for all amino acids comprising 90% identity to SEQ ID NO: 79, 90 and 91. As noted above, it is assumed for the sake of compact prosecution that these sequences refer to Cas proteins, rather than adenosine deaminase. The specification as filed does not provide sufficient evidence that Applicants were in possession of the full scope of the claimed invention at the time of filing of the instant invention. As such, >90% identity to a 1404 amino acid polynucleotide, polynucleotide encompasses
1404
C
140
×
3
140
=
m
o
r
e
t
h
a
n
15
×
10
262
amino acid molecules, considering substitution mutations (i.e. not including insertions and deletions). This amounts to enormous number of polynucleotides which do not have support in the specification. The specification also does not provide any guidance on how or where the amino acids need to be changed. It is clear that the vector of the invention was restricted to SEQ ID NOs: 79, 90 and 91. For example, see instant specification at [0153] – [0159] and [0165].
Similar rejections apply to claim 114.
Claims 2, 6-7, 10, 14, 16, 18, 22, 23, 26, 28, 32-37, 76 and 114 are rejected for their dependency on above rejected claims.
Regarding claim 76: The claim states that:
“The method of claim 1, wherein the step of contacting results in a sequence that is not associated with Usher syndrome an increase in full-length Usherin protein, partial or complete restoration of Usherin protein stability or function, partial or complete reversal of visual or hearing loss, or partial or complete restoration of auditory or visual function.”
The Applicants have claimed a method of completely or partially treating visual or hearing loss, or partial or complete restoration of auditory or visual function using their adenosine-Cas9 editing method. However, the experimental data in the application did not teach partial or complete reversal of visual or hearing loss, or partial or complete restoration of auditory or visual function. Therefore, at most Applicant’s data taught A to G editing in usherin protein in humans’ cells (HEK293) and Rhesus monkey cells. None of the experiments taught teach partial or complete reversal of visual or hearing loss, or partial or complete restoration of auditory or visual function using the claimed method of claim 1. It is also pointed out that the specification also did not teach complete or partial restoration of Usherin protein stability or function.
“An original claim may lack written description support when (1) the claim defines the invention in functional language specifying a desired result but the disclosure fails to sufficiently identify how the function is performed or the result is achieved…” (See MPEP 2163.03 V). MPEP §2163.03(a) makes clear that functional claim limitations must be supported by an adequate written description showing possession of the claimed invention. Here, the specification lacks any teaching, guidance, or evidence enabling a POSITA to conclude that performing the claimed method would achieve the functional outcomes recited. The claim is thus overbroad, attempting to claim results that extend beyond what the inventors actually possessed.
Furthermore, without demonstration or disclosure of how base editing translates to functional protein restoration or phenotypic improvement, the claim fails to convey that the inventors were in possession of the invention as claimed. Consequently, the claim does not meet the requirements of §112(a) for written description.
Claim 76 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement.
The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
Applicant's specification is not found to be enabling for method of complete or partial restoration of Usherin protein stability or function or a method of completely or partially treating visual or hearing loss, or partial or complete restoration of auditory or visual function using their adenosine-Cas9 editing method.
Analysis of whether a particular claim is supported by the disclosure in an application requires a determination of whether that disclosure, when filed, contained sufficient information regarding the subject matter of the claims as to enable one skilled in the pertinent art to make and use the claimed invention without undue or unreasonable experimentation. See Mineral Separation v. Hyde, 242 U.S. 261, 270 (1916). The key word is 'undue,' not experimentation.' " (Wands, 8 USPQ2d 1404). The factors to be considered in determining whether undue experimentation is required are summarized In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988). The factors to be considered in determining whether undue experimentation is required include: (1) the quantity of experimentation necessary, (2) the amount or direction or guidance presented, (3) the presence or absence of working examples, (4) the nature of the invention, (5) the state of the prior art, (6) the relative skill of those in the art, (7) the predictability or unpredictability of the art, and (8) the breadth of the claims. While all these factors are considered, a sufficient number are discussed below so as to create a prima facie case.
Applicants' claims are directed to a method of complete or partial restoration of Usherin protein stability or function or a method of completely or partially treating visual or hearing loss, or partial or complete restoration of auditory or visual function using their adenosine-Cas9 editing method.
The specification did not provide evidence for partial or complete restoration of protein stability or function or partial or complete reversal off hearing or visual loos using their claimed method.
At the time the invention was made it was known that therapeutic rescue of Usher-syndrome phenotypes was highly uncertain, even when gene expression could be restored. In Isgrig et al. (See PTO-892), the authors reported partial hearing rescue in a different Usher subtype (USH1G) following AAV-mediated gene supplementation (Isgrig, Nat. Biotechnol. 2017/2018). Isgrig emphasized the incomplete and variable restoration of hearing (See Isgrig et al p. 784, col. 1, last para). Further as evidenced by Mauriac, “Cochlear implants or hearing aids are the standard of care for USH2A hearing impairment, but no biological treatments are available that alter the progression of the disease, which leads to vision loss in USH2A patients. “ (See Mauriac, p. 2391, col. 2, last para). Mauriac further taught that role of USH2A in retina is poorly understood. (See Mauriac, p. 2391, col. 2, first para). As such the prior art did not teach partial or complete restoration of hearing loss or vision loss was possible by editing G to A mutations.
Therefore, there was a recognized level of unpredictability with regards to underlying molecular mechanisms involved in treatment of usher syndrome. Due to the lack of teachings in the art regarding treatment of usher syndrome, and the recognized unpredictability in ameliorating the symptoms, a large amount of guidance and teachings would be necessary in order to be enabling for methods of such.
Guidance and teachings provided by Applicants in the instant specification is limited to disclosure about plausibility of base editing of USH2A in vitro. For example, [0543] taught that it was possible to introduce skipping of exons by mutating splice sites of the Ush2a gene in mouse organ of Corti (OC-k1) cells and mouse retina. (See Figure 10). Further the specification provided guidance about in vitro editing of human cells and rhesus monkey cells. It is acknowledged that the Office does not require the presence of working examples to be present in the disclosure of the invention (see MPEP §2164.02). However, in light of the state of the art, discussed above, which recognizes a high level of unpredictability in the field of treatments for cardiac ischemia, and teachings of uncertainty regarding genetic correlation causing symptoms of Usher syndrome, with no consensus being reached using any single protein for treatment of Usher syndrome, the Office would require appropriate disclosure to support the contention that method of claim 1 may be successfully employed in fully or partially resolving hearing or vision loss in usher syndrome. The amount of guidance or direction needed to enable the invention is inversely related to the amount of knowledge in the state of the art as well as the predictability in the art. In re Fisher, 427 F.2d 833, 839, 166 USPQ 18, 24 (CCPA 1970). Thus, due to the high level of unpredictability in the art, the current specification would have to provide greater amounts of teachings and guidance directed to methods of carrying out the claimed invention.
Therefore, due to the sum of all the aforementioned factors, one of ordinary skill in the art, at the time the invention was made, would not expect success carrying out the claimed method of claim 76. Given that the art fails to recognize and Applicant has failed to demonstrate that the claimed method would alleviate symptoms of ischemic heart disease, the skilled artisan would be faced with the impermissible burden of undue experimentation in order to practice the claimed invention. Accordingly, claim 76 is deemed properly rejected.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
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 1-2, 6, 10, 14, 16 and 32-35 are rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 04/14/2023) and WO2016005514A1 (Published Jan 14, 2016; See PTO-892).
Regarding claims 1-2, 6, and 14: Fry taught the importance of deaminating genes like USH2A to correct G[Wingdings font/0xE0]A and T[Wingdings font/0xE0]C mutations in RNA as a therapeutic for genetic diseases. (See Fry Abstract). Fry further taught that “[s]ite-directed RNA editing uses molecular tools to recruit RNA editing enzymes to target sites of interest and enables the targeted correction of G>A and T>C mutations in coding sequences of RNA.” (See Fry, p. 3, para 1). Fry explained that “The majority of strategies to edit RNA utilize the activity of the naturally occurring human adenosine deaminase acting on RNA (ADAR) enzymes. ADARs act to convert adenosine residues to inosines (A>I editing) in double-stranded RNA. Inosine is structurally similar to guanosine and is read as a guanosine by most cellular machinery including during translation, splicing and reverse transcription, effectively creating an A>G edit in RNA.” (See Fry, Fig. 2; p. 3, para 3). Fry further explained that “the hyperactive ADAR2DD(E488Q) can produce numerous off-target editing events, so further rational mutagenesis was undertaken to generate a more specific ADAR2DD(E488Q/T375G) deaminase fused with dPspCas13b (termed REPAIRv2).” The ADAR2DD(E488Q/T375G) deaminase reads on the claimed base editor.
Fry did not teach a DNA binding protein, as a targeting nuclease. Yang taught that “[m]ore recently, adenine base editors (ABEs), which efficiently convert A•T base pairs to G•C in genomic DNA, have been developed via fusion of an engineered tRNA adenosine deaminase (ecTadA from Escherichia coli) with nCas9” (See Yang p. 814 col. 1, first para). Yang further demonstrated “an increase of ABE activity through injection of chemically modified tracrRNA and crRNA in mouse zygotes, and the expansion of editing scope by fusion of an ecTadA (as required by claim 6) mutant to SaCas9n-KKH (as required by claim 14) and Cas9n-VQR variants in both cells and embryos.” (See Yang p. 814 col. 1, first para). It would have been obvious for a person of ordinary skill in the art to replace the deaminase domain and nuclease domain taught by Fry with the domain taught by Yang to acieve deamination of genes such as USH2A. The person would recognize this as simple substitution useful for the same purpose ((KSR Int’l Co. v. Teleflex, Inc., 550 U.S. 398 (2007) pg 14 and 12). The Cas9 as taught by Yang reads on the claimed napDNAbp.
It is recognized that Fry did not teach a nucleic acid having at least 90% identity to any one of SEQ ID NOs: 12-35. However, it would have been obvious for a person of ordinary skill in the art to arrive at any one or all of the claimed sequences as the sequence of USH2A is well known. See WO2016005514A1 SEQ ID NO: 1. Further Fry taught a method of developing guides for deaminating with mismatches, “Guides with homology regions of between 30 and 84 nucleotides and with an A-C mismatch to the target adenosine placed across a range of positions within the homology region were found to be functional. However, 50 nucleotide guides with a mismatch placed 32–36 nucleotides from the 50 end was found to be generally most efficacious. Fry also taught a c.11864G>A mutation in usherin gene (as required by claim 16) result in a codon change from tryptophan (TGG) to a premature termination codon (TAG). (See Fry p. 13, 2nd para). As such it would have been obvious for a person of ordinary skill in the art to determine a guide sequence based on the teachings of Fry. The person will be motivated to use base editor complexed with Cas protein for deaminating common genetic mutations of a gene, such as Ush2A for treatment of genetic diseases.
Fry teaches base editing of USH2A using adenosine deaminases and provides detailed guidance for designing functional gRNAs, including length, mismatch placement, and homology regions. Yang teaches a TadA-Cas9 fusion capable of targeted A→G base editing in genomic DNA and broadening the editing scope through Cas9 variants. WO2016005514A1 discloses the USH2A sequence, enabling a POSITA to select complementary target sequences. It would have been predictable and routine for a person of ordinary skill in the art to combine Fry’s gRNA design principles with Yang’s TadA-Cas9 base editor to target adenosines in USH2A, including sequences corresponding to SEQ ID NOs: 12–35 or naturally occurring variants, for performing the claimed method. Therefore, the claimed method would have been obvious to a POSITA at the time of invention.
Regarding claim 10: Yang taught “ABE and its variants efficiently generate site-specific A:T>G:C conversions in cell lines, mouse and rat embryos.” (See Yang p. 818, col. 1, para 2). Yang used ABE7.10 for A>G conversion in rodent embryos. (See Yang p. 815, col. 1, line 2).
Regarding claim 16: Fry taught a G11864A which results in a codon change from tryptophan to a premature termination codon as a common USH2A mutation. (See Fry p. 13, para 2).
Regarding claim 32-35: Fry taught “an approach to RNA editing in a patient with Usher Syndrome type 2 with compound heterozygous mutations in USH2A (c.2299delG / c.11864G>A).” (See Fry Figure 5). Fry further explicitly taught that “the growing armament of tools for RNA editing will allow the interrogation of a range of approaches for safety and efficacy in vivo.” (See Fry p. 15, last para). It is submitted that Fry clearly anticipated in vivo delivery such as to the inner ear for Usher syndrome in human subjects.
Claim 7 is rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 04/14/2023); WO2016005514A1 (Published Jan 14, 2016; See PTO-892) and WO2016022363A2 (Published Feb 11, 2016; See IDS of 4/14/2023).
Regarding claim 7: The teachings of Fry, in view of Yang and WO2016005514A1 are set forth above. As explained above, for the sake of compact prosecution, it is assumed that the Applicants claim a Cas9 protein that is at least 90% identical to the claimed SEQ ID NOs: 79, 90-91. WO2016022363A2 taught a Cas9 protein comprising a SEQ ID NO: 150 which is 100% identical to instant SEQ ID NO: 79 (See alignment below) (See specification of WO2016022363A2 [00152]). As such it would have been obvious to use the Cas9 taught by WO2016022363A2 in the construct taught by Yang. It would have been obvious to a POSITA to substitute the Cas9 of Fry/Yang with the Cas9 taught in WO2016022363 because these enzymes are known, functionally equivalent, and routinely substituted in the art. (See WO2016022363A2 [00169]).
Therefore, an Cas9 domain having at least 90% identity to Applicant’s SEQ ID NOs: 79, 90, or 91 would have been an obvious variant in view of the combined teachings of the references.
Query Match 100.0%; Score 7206; Length 1404;
Best Local Similarity 100.0%;
Matches 1404; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAE 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAE 60
Qy 61 ATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG 120
Qy 121 NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD 180
Qy 181 VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGN 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGN 240
Qy 241 LIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 LIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI 300
Qy 301 LLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 LLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA 360
Qy 361 GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELH 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELH 420
Qy 421 AILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEE 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 AILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEE 480
Qy 481 VVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFL 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 VVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFL 540
Qy 541 SGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKI 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 SGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKI 600
Qy 601 IKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 IKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG 660
Qy 661 RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL 720
Qy 721 HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER 780
Qy 781 MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDH 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDH 840
Qy 841 IVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNL 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 IVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNL 900
Qy 901 TKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKS 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKS 960
Qy 961 KLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 KLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 1020
Qy 1021 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF 1080
Qy 1081 ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVA 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVA 1140
Qy 1141 YSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPK 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 YSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPK 1200
Qy 1201 YSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVE 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 YSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVE 1260
Qy 1261 QHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 QHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 1320
Qy 1321 PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVSSD 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVSSD 1380
Qy 1381 YKDHDGDYKDHDIDYKDDDDKAAG 1404
||||||||||||||||||||||||
Db 1381 YKDHDGDYKDHDIDYKDDDDKAAG 1404
Alignment between SEQ ID NO: 150 of WO2016022363A2 and instant SEQ ID NO: 79.
Claims 18, 26 and 28 are rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 4/14/2023); WO2016005514A1 (Published Jan 14, 2016; See PTO-892) and WO2015079057A2 (Published Jun 04, 2015; See PTO-892).
Regarding claim 26: The teachings of Fry in view of Yang and WO2016005514A1 are set forth above. However, the cited references did not teach an gRNA that is 85% identical to SEQ ID NO: 170. However, WO2015079057A2 taught a SEQ ID NO: 4, a “constant part of the gRNA, including the constant part of crRNA, a linker and the tracrRNA” (See WO2015079057A2 ) which is 100% identical to instant SEQ ID NO: 170. (See alignment below)
Query Match 100.0%; Score 76; Length 76;
Best Local Similarity 100.0%;
Matches 76; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGT 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGT 60
Qy 61 GGCACCGAGTCGGTGC 76
||||||||||||||||
Db 61 GGCACCGAGTCGGTGC 76
Alignment between SEQ ID NO: 4 of WO2015079057A2 with instant SEQ ID NO: 170.
As SEQ ID NO: 170 represents a known, widely used canonical gRNA constant scaffold, a person of ordinary skill in the art would have been motivated to use this same constant gRNA sequence in the base-editing compositions taught by Fry and Yang to achieve predictable and routine CRISPR guide function. Substituting a known, gRNA scaffold (WO2015079057A2 SEQ ID NO:4) for the corresponding guide RNA of Fry or Yang would have been a simple, predictable substitution yielding a gRNA having at least 85% identity to SEQ ID NO:170, with a reasonable expectation of success. Accordingly, the claimed gRNA would have been obvious.
Regarding claims 18 and 28: As pointed out above, A person of ordinary skill in the art would have been motivated to combine Fry, Yang, and WO2016005514A1 because Yang’s improved ABE constructs were widely recognized as providing increased efficiency and fidelity, and WO2016005514A1 provides known, validated USH2A sequence information necessary for implementing Fry’s editing strategy. Such a combination represents routine, predictable optimization of known components in the base-editing field. It is also noted that Yang taught that adenosine base editors are guided to genomic targets through gRNAs containing standard ~20-nucleotide spacer sequences complementary to the genomic protospacer region. Yang therefore provides the conventional gRNA architecture including the 20-nt spacer by which a POSITA would implement an ABE at any selected genomic target.
WO2015079057 further teaches a crRNA–linker–tracrRNA guide RNA architecture in which SEQ ID NO:4 (which is identical to applicant’s SEQ ID NO:170) is expressly disclosed as the constant gRNA region used in constructing such CRISPR guides. A POSITA would have recognized that, to implement Fry’s USH2A targeting with the ABE system of Yang, it would have been routine to combine a selected 20-nt spacer sequence (such as those recited in the claim, e.g., SEQ ID NOs: 36, 46, 47, 50, etc.) with a standard CRISPR constant region, such as SEQ ID NO: 170, because CRISPR systems universally require a conserved structural region paired with a variable spacer.
Thus, in view of Fry’s identification of the USH2A target region, Yang’s teaching of standard CRISPR-based ABE gRNA architecture and 20-nt spacers, WO2016005514’s disclosure of the USH2A sequence enabling routine spacer design, and WO2015079057’s explicit disclosure of the constant region corresponding to applicant’s SEQ ID NO:170, a POSITA would have been motivated to construct a gRNA comprising a ~20-nt spacer to target Fry’s USH2A site and the known constant region of WO2015079057, with a reasonable expectation of success. Accordingly, the claimed gRNA containing the recited spacer and the second nucleic acid comprising SEQ ID NO: 170 would have been obvious.
Claims 22-23 are rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 4/14/2023); WO2016005514A1 (Published Jan 14, 2016; See PTO-892); and WO2015079057A2 (Published Jun 04, 2015; See PTO-892) as applied to claim 18 and further in view of Winter et al (Published 2019; hereinafter "Winter;" See IDS of 4/14/2023) Pendse et al (Published 2020.02.04; Cold Spring Harbor Laboratory; See IDS of 4/14/2023).
Regarding claim 22-23: The teachings of Fry, in view of Yang and WO2016005514A1 are set forth above. Briefly, Fry taught adenosine base editing of the USH2A gene and discloses guide RNA design parameters, including homology lengths and mismatch positioning, that directly correspond to the claimed guides having at least 90% identity to SEQ ID NOs: 12–35. Fry therefore provides both the target gene (USH2A) and the use of adenosine deamination guided by sequence-complementary gRNAs. Yang taught the canonical ABE architecture comprising a nucleic-acid-programmable DNA-binding protein (e.g., Cas9) fused to an adenosine deaminase (TadA), thereby supplying the well-known structural configuration of the base editor used in Fry’s system. WO2016005514A1 further taught the USH2A genomic sequence and protospacer regions corresponding to the claimed guide sequences. A person of ordinary skill in the art would have been motivated to combine Fry, Yang, and WO2016005514A1 because Yang’s improved ABE constructs were widely recognized as providing increased efficiency and fidelity, and WO2016005514A1 provides known, validated USH2A sequence information necessary for implementing Fry’s editing strategy. Such a combination represents routine, predictable optimization of known components in the base-editing field.
However, none of the cited references taught use of the construct for exon skipping in USH2A genes. However, Winter taught that “A > G base editors (ABE) to induce exon skipping by mutating the adenine in the highly conserved AG dinucleotide within splice acceptors.” (See Winter, p.2, col. 1, second para). Further Pendse taught that “skipping of exon 13 of the human USH2A gene or the equivalent exon 12 of the mouse Ush2a gene results in an in-frame transcript that produces functional Usherin protein.” (See Pendse Abstract, as required by claim 23). As such Winter further taught that adenosine base editors are suitable for inducing exon skipping by mutating the adenine of the highly conserved AG splice-acceptor motif. Winter therefore provided the precise editing mechanism recited in the claims ABE-mediated disruption of a splice acceptor to force exon removal. Pendse teaches that skipping exon 13 of USH2A yields an in-frame transcript that produces functional usherin, thereby providing a direct therapeutic motivation to induce exon skipping at this exon. A person of ordinary skill in the art would have been motivated to apply Winter’s ABE-mediated splice-acceptor-editing technique to the exon-13 splice acceptor of USH2A in view of Pendse’s explicit teaching that exon 13 skipping is beneficial and preserves protein function. Accordingly, it would have been obvious to employ the Fry/Yang/WO2016005514A1 base-editing system to disrupt the exon-13 splice acceptor as taught by Winter, motivated by the functional outcomes disclosed in Pendse, yielding the claimed method with predictable results.
Claims 36-37 are rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 4/14/2023); WO2016005514A1 (Published Jan 14, 2016; See PTO-892) and Richter et al (Nat Biotechnol. 2020 Jul; hereinafter "Richter;" See IDS 11/07/2025).
Regarding claims 36-37: The teachings of Fry in view of Yang and WO2016005514A1 are set forth above. None of the cited references disclose or teach ABE8e as the base editor. However, Richter disclosed that they “evolved the deaminase component of ABE7.10 using phage-assisted non-continuous and continuous evolution (PANCE and PACE), which resulted in ABE8e. ABE8e contains eight additional mutations that increase activity (kapp) 590-fold compared with that of ABE7.10. ABE8e offers substantially improved editing efficiencies when paired with a variety of Cas9 or Cas12 homologs.” (See Richter Abstract).
A person of ordinary skill in the art seeking to improve editing efficiency of Fry’s USH2A-targetting methods would have been motivated to substitute the ABE8e/Cas system taught by Richter, particularly as Richter taught higher editing efficiency and predictable outcomes. Accordingly, in view of Fry, Yang, WO2016005514A1, and Richter, claims 36-37 are obvious.
Claim 114 is rejected under 35 U.S.C. 103 as being unpatentable over Fry et al (Int J Mol Sci. 2020 Jan 25; hereinafter "Fry;" See PTO-892) in view of Yang et al (Protein Cell. 2018 Sep; hereinafter "Yang;" See IDS of 4/14/2023); WO2016005514A1 (Published Jan 14, 2016; See PTO-892) and US10113163B2 (Published Feb 08, 2018; IDS of 4/14/2023).
Regarding claim 114: The teachings of Fry in view of Yang and WO2016005514A1 are set forth above. None of the cited references disclose or teach a base editor having at least 95% identity to SEQ ID NOs: 373-388. However, US10113163B2 disclosed a base editor-napDNAbp comprising SEQ ID NO: 11, which has 100% identity to instant SEQ ID NO: 373. (See alignment below). (See corresponding PCT claim 169).
Query Match 100.0%; Score 8011; Length 1561;
Best Local Similarity 100.0%;
Matches 1561; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI 60
Qy 61 MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDV 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDV 120
Qy 121 LHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGSETPGTSESAT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 LHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGSETPGTSESAT 180
Qy 181 PESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 PESDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET 240
Qy 241 AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI 300
Qy 301 FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN 360
Qy 361 SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF 420
Qy 421 GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSD 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSD 480
Qy 481 AILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNG 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 AILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNG 540
Qy 541 YAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 YAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGE 600
Qy 601 LHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNF 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 LHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNF 660
Qy 661 EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 EEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA 720
Qy 721 FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLL 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLL 780
Qy 781 KIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 KIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTG 840
Qy 841 WGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 WGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD 900
Qy 901 SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSR 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSR 960
Qy 961 ERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 ERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV 1020
Qy 1021 DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFD 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFD 1080
Qy 1081 NLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITL 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 NLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITL 1140
Qy 1141 KSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDV 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 KSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDV 1200
Qy 1201 RKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 RKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR 1260
Qy 1261 DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT 1320
Qy 1321 VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKL 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKL 1380
Qy 1381 PKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLF 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1381 PKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLF 1440
Qy 1441 VEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNL 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1441 VEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNL 1500
Qy 1501 GAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRK 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 GAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRK 1560
Qy 1561 V 1561
|
Db 1561 V 1561
As SEQ ID NO: 373 represents a known, widely used adenosine base editor, a person of ordinary skill in the art would have been motivated to use this same ABE in the base-editing compositions taught by Fry and Yang to achieve predictable and routine CRISPR guide and base editing function. Substituting a known ABE (US10113163B2 SEQ ID NO: 11) for the base editor Fry or Yang would have been a simple, predictable substitution, with a reasonable expectation of success. Accordingly, the claimed base editors would have been obvious.
Conclusion
No claim is free of art.
No claim is allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAGAMYA VIJAYARAGHAVAN whose telephone number is (703)756-5934. The examiner can normally be reached 9:00a-5:00p.
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