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 .
Claims Status
Applicant’s response filed 10/31/2025 has been received and considered entered. This is a response to amendments and arguments filed 10/31/2025.
Claim 8 stands withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected inventions, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 01/08/2025.
Claims 3-7, 9, 18, 20, 26 is/are cancelled. Claims 1-2, 8, 10-17, 19, 21-25 is/are currently pending with claim 8 withdrawn. Claims 1-2, 10-17, 19, 21-25 is/are under examination.
Information Disclosure Statement
The listing of references in the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892, they have not been considered.
The information disclosure statement filed 10/31/2025 fails to comply with 37 CFR 1.98(a)(1), which requires the following: (1) a list of all patents, publications, applications, or other information submitted for consideration by the Office; (2) U.S. patents and U.S. patent application publications listed in a section separately from citations of other documents; (3) the application number of the application in which the information disclosure statement is being submitted on each page of the list; (4) a column that provides a blank space next to each document to be considered, for the examiner’s initials; and (5) a heading that clearly indicates that the list is an information disclosure statement. The information disclosure statement has been placed in the application file, and the information referred to therein has been considered; however, the following document provided by the applicant has not been listed in the IDS. Applicant has cited the NPL “How to Choose the Right Cas Variant for Every CRISPR Experiment” in the provided arguments (page 11), and has provided a copy. However, this document was not cited in the provided IDS.
Claim Interpretation
Claims 1 and 19 recite that the myristoylation motif is followed by positively-charged amino acids. However, claim 19 does not recite whether these positively-charged amino acids must be consecutive or must immediately follow the myristoylation motif. As such, this limitation is interpreted to encompass positively-charged amino acids which are downstream of the myristoylation motif but are not immediately downstream of the myristoylation motif.
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.
Claim(s) 1-2, 12-15, 19 is/are rejected under 35 U.S.C. 103 as being unpatentable over Dominguez-Monedero (2018), and further in view of Campbell (January 2019), Hayashi (2010), and Shen (2011). This rejection is maintained, with amendments highlighting teachings from the references which had not previously been discussed.
Regarding claim 1, Dominguez-Monedero teaches a fusion protein comprising an N-terminal myristoylation domain, a Cas9 domain, and a nuclear localization signal (Figure 5; pages 2164-2165), wherein the myristoylation domain does not comprise a palmitoylation motif (Supplementary Table 1). Dominguez-Monedero teaches that the myristoylation domain comprises the amino acid sequence GSSKSKPKDPSQR (i.e., myristoylation motif G-X1-X1-X1-S/T followed by at least one positively-charged amino acid, underlined) (Supplementary Table 1; amino acid sequence was determined using known codon translations). Dominguez-Monedero additionally teaches Myr-Cas9 fusion proteins and Cas9-NLS fusion proteins (i.e., fusion proteins consisting of a myristoylation domain and a Cas9 domain or consisting of a Cas9 domain and an NLS domain) (page 2167). Dominguez-Monedero further teaches that the function of the myristoylation domain is targeting of the fusion protein to the plasma membrane (page 2164), and the function of the NLS domain is targeting of the fusion protein to the nucleus (page 2164).
Regarding claim 2, Dominguez-Monedero teaches that the myristoylation domain comprises the amino acid sequence GSSKSKPKDPSQR (i.e., G-X1-X1-X1-S/T-X2-X2-X2) (Supplementary Table 1; amino acid sequence was determined using known codon translations).
Regarding claim 12, Dominguez-Monedero teaches that the Cas9 domain is a wildtype Cas9 (Fig. 5)
Regarding claim 13, Dominguez-Monedero teaches that dCas9 (nuclease dead Cas9) has been used in the art for applications such as epigenetic regulation (page 2162). It would have been obvious to a person of ordinary skill in the art at the time of filing that the wild-type Cas9 of the Myr-hPR-Cas9-NLS or Myr-Cas9-ERT2-NLS-LEP (page 2167) constructs of Dominguez-Monedero could be replaced with dCas9 in applications such as epigenetic regulation, where DNA cleavage would not be required.
Regarding claims 14 and 15, Dominguez-Monedero teaches that the NLS comprises the amino acid sequence PKKKRKV and is an SV40 NLS (Supplementary Table 1; page 2167).
However, while Dominguez-Monedero does teach a myristoylation domain wherein the full sequence of the myristoylation domain of Dominguez-Monedero exceeds 10 amino acids.
Hayashi teaches that myristoylation domains which are longer than 10 amino acids long can be truncated to consist of 5-10 amino acids.
Regarding claims 1-2, Hayashi teaches that the myristoylation domain taught by Dominguez-Monedero (GSSKSKPKDPSQR) comprises a shorter, functional myristoylation domain (GSSKSKPKDP), which comprises the myristoylation motif GSSKS followed by at least one positively-charged amino acid (K, underlined) (Table 3).
It would have been obvious to a person of ordinary skill in the art at the time of filing that any known myristoylation motif which targets a protein to the plasma membrane could be substituted for the myristoylation motif taught by Dominguez-Monedero, including those taught by Hayashi. Furthermore, as discussed above, Hayashi teaches that the core functional unit of a myristoylation domain is 10 amino acids long (see Table 3, particularly the last row, “Residues required for the CaM binding: G-KLS-----“; page 495, “the enzyme recognizes approximately only ten residues from the N-termini of substrates, and there is no consensus sequence for myristoylation besides glycine at the second position and serine at the sixth position, i.e., MGXXXSXX in the precursor proteins”). It would have been obvious to an artisan at the time of filing that the minimal length of a myristoylation domain—10 amino acids—was equivalent in function, as a myristoylation domain, to a longer sequence comprising the same core domain.
However, Dominguez-Monedero and Hayashi do not teach that Cas9 and gRNA can be packaged as a ribonucleoprotein complex in an extracellular vesicle.
Regarding claim 1, Campbell teaches that Cas9 and gRNA can be delivered as a ribonucleoprotein complex encapsulated in an extracellular vesicle (abstract; “Introduction” section pages 151-152) (required for claims 6-7).
Campbell teaches that Cas9 proteins can be delivered to cells as a ribonucleoprotein complex (i.e. the Cas9 protein conjugated with a gRNA) encapsulated in an extracellular vesicle. While Dominguez-Monedero solely teaches introducing Cas9 and gRNAs into cells as a polynucleotide encoding the Cas9 fusion protein and gRNA, the teachings of Campbell make clear that at the time of filing, it was known in the art that Cas9 and gRNA could alternatively be introduced into cells as a ribonucleoprotein complex encapsulated in an extracellular vesicle. Campbell teaches that extracellular vesicles are ideal vehicles for encapsulation and delivery of Cas9/gRNA ribonucleoprotein complexes, as extracellular vesicles are used and created endogenously by cells, and thus can be readily formed and taken up by cells with reduced toxicity compared to plasmid-based delivery mechanisms (pages 151-152, 156). Based on the teachings of Campbell, it would have been obvious to a person of ordinary skill in the art at the time of filing that the fusion Cas9 protein and gRNA of Dominguez-Monedero should be encapsulated in an extracellular vesicle as a ribonucleoprotein complex to both reduce cytotoxicity from plasmid-based delivery of Cas9/gRNA and improve the capacity of the Cas9/gRNA complex being taken up by target cells, as Campbell teaches these as beneficial features of extracellular vesicle encapsulation of Cas9/gRNA complexes as compared to plasmid-based introduction of Cas9/gRNA systems into cells.
While Campbell teaches a mechanism for encapsulation in an extracellular vesicle which would require additional domains added to the myr-Cas9-NLS fusion protein rendered obvious by Dominguez-Monedero and Hayashi, Shen teaches N-terminal myristoylation domains that are 5-10 amino acids long target a fusion protein to be encapsulated in an extracellular vesicle.
Regarding claim 1, Shen teaches an N-terminal myristoylation domain that is 5-10 amino acids in length (page 14385: MGAINSKRKD). Shen further teaches that a myristoylation motif lacking a palmitoylation motif can effectively target a protein to be encapsulated into an extracellular vesicle (page 14385). This myristoylation domain comprises at least one positively-charged amino acid following the myristoylation motif and lacks a palmitoylation motif (MGAINSKRKD).
Regarding claim 2, Shen teaches the myristoylation domain MGAINSKRKD (page 14385). Hayashi teaches that the N-terminal methionine is removed by methionyl aminopeptidase, exposing the glycine residue immediately C-terminal of this methionine, as required for myristoylation (page 495). As such, it would be obvious to an artisan that in a cell, the myristoylation motif of Shen would be modified by methionyl aminopeptidase to GAINSKRKD, which comprises the general motif G-X1-X1-X1-S/T, as required by claim 2.
Dominguez-Monedero teaches that the purpose of the myristoylation domain is to facilitate binding of the fusion protein to the cell membrane. As such, it is the myristoylation modification which Dominguez-Monedero specifies as necessary, and not necessarily the particular myristoylation motif sequence. As such, it would have been obvious to a person of ordinary skill in the art at the time of filing that any myristoylation motif which facilitates cell/plasma membrane targeting (including that taught by Shen) could be substituted for the myristoylation motif of Dominguez-Monedero, while maintaining the expressed purpose of the added myristoylation motif. As the myristoylation motif of Shen targets proteins to the cell membrane (page 14385), an artisan would find obvious that the myristoylation motif of Shen could be substituted for the myristoylation of Dominguez-Monedero.
Regarding claim 1, while Dominguez-Monedero does not teach fusion proteins that only comprise an N-terminal myristoylation domain, a Cas9 domain, and an NLS, with no other structures, Dominguez-Monedero, combined with Campbell, Shen, and Hayashi, renders obvious fusion proteins consisting of an N-terminal myristoylation domain, a Cas9 domain, and an NLS. The additional hPR, ERT2, or LEP domains and the myristoylation domain taught by Dominguez-Monedero are not considered to materially affect the basic and novel characteristic(s) of the claimed invention. While the stated novelty of Dominguez-Monedero is the drug-inducible activation of Cas9 function, an artisan would find obvious that Dominguez-Monedero additionally teaches Cas9 fusion proteins having cell membrane-targeting capacity, through a myristoylation domain, and nucleus-targeting capacity, through an NLS domain. The claimed invention requires that the ribonucleoprotein complex of the fusion protein and a guide RNA be encapsulated in an extracellular vesicle (described above by Campbell). Based on the above-described combination of Dominguez-Monedero and Campbell, a Cas9 ribonucleoprotein complex, including one wherein the Cas9 is a fusion protein according to Dominguez-Monedero, can be encapsulated in an extracellular vesicle. The instant specification discloses that the presence of a particular myristoylation motif facilitates encapsulation in extracellular vesicles in specific cells (see paragraphs [0110], [0149]-[0150]: PC3, DU145, LNCaP, and 22Rv1 cells; see also Figs. 17A-E). Applicant has argued in the response filed 07/02/2025 that the novelty of the claimed invention lies in the use of a myristoylation signal to encapsulate the fused protein in an extracellular vesicle. However, the art, as exemplified in Shen, teaches that myristoylation motifs were already known in the art to facilitate encapsulation in extracellular vesicles as an inherent result of myristoylation of the proteins (Shen page 14385). Given the claimed novelty of the claimed invention as argued by the applicant in the response filed 03/28/2025, based on the teachings of the prior art, the claimed novelty and basic characteristics of the claimed invention would not materially be affected by additional structural components of Dominguez-Monedero. Dominguez-Monedero teaches that the addition of an N-terminal myristoylation motif and additional domains does not prevent the function of a fused Cas9 protein and Shen teaches that myristoylation can be used to encapsulate proteins in extracellular vesicles.
Claim(s) 16-17 and 22 is/are rejected under 35 U.S.C. 103 as being unpatentable over Dominguez-Monedero (2018), Campbell (January 2019), Hayashi (2010), and Shen (2011) as applied to claim 1 above, and further in view of Chiper (2017). This rejection is maintained.
The teachings of Dominguez-Monedero, Campbell, Hayashi, and Shen are discussed above and render obvious the limitations of claim 1. However, they do not teach the nuclear localization signals claimed in claims 16-17 and 22.
Chiper teaches NLS sequences.
Regarding claims 16-17 and 22, Chiper teaches that fusion proteins can comprise nuclear localization signals of many different origins, including SV40 large T antigen (PKKKRKV), c-Myc (PAAKRVKLD), nucleoplasmin (AVKRPAATKKAGQAKKKKLD), and Tus protein (KLKIKRPVK) (page 1701040).
Chiper teaches that different NLS have different functional qualities. For example, Chiper teaches that the c-Myc NLS allows for greater nuclear accumulation of the fused protein than the SV40 NLS (page 1701040). Dominguez-Monedero teaches that the fusion protein comprises an SV40 NLS (Supplementary Table 1; page 2167). Given the teachings of Chiper, it would have been obvious to a person of ordinary skill in the art at the time of filing that the SV40 NLS used by Dominguez-Monedero should be substituted with the c-myc NLS, as taught by Chiper, in order to increase the rate of fusion protein localization to the nucleus.
Claim(s) 16-17 and 21-22 is/are rejected under 35 U.S.C. 103 as being unpatentable over Dominguez-Monedero (2018), Campbell (January 2019), Hayashi (2010), and Shen (2011) as applied to claim 1 above, and further in view of Ray (2015). This rejection is maintained.
The teachings of Dominguez-Monedero, Campbell, Hayashi, and Shen are discussed above and render obvious the limitations of claim 1. However, these references do not teach that the NLS sequence is derived from nucleoplasmin, EGL-13, c-Myc, or TUS-protein.
Ray teaches the NLS sequences of EGL-13, c-Myc, NLP, and TUS.
Regarding claim 16, Ray teaches the NLS sequences of EGL-13, c-Myc, NLP, and TUS.
Regarding claim 17, Ray teaches the NLP NLS sequence AVKRPAATKKAGQAKKKKLD (Fig. 1).
Regarding claim 21, Ray teaches that the EGL-13 NLS sequence is MSRRRKANPTKLSENAKKLAKEVEN (Fig. 1).
Regarding claim 22, Ray teaches the NLS sequences PAAKRVKLD and KLKIKRPVK (Fig. 1).
Ray teaches that the NLS sequences derived from EGL-13, SV40, c-Myc, NLP, and TUS exhibit different levels of nuclear targeting when fused to a protein (Fig. 1). As such, it would have been obvious to a person of ordinary skill in the art at the time of filing to modify the fusion protein rendered obvious by Dominguez-Monedero, Campbell, and Shen with any one of the NLS sequences taught by Ray, thereby enabling the artisan to modulate the strength of targeting of the fusion protein to the nucleus.
Claim(s) 24-25 is/are rejected under 35 U.S.C. 103 as being unpatentable over Dominguez-Monedero (2018), Campbell (January 2019), Hayashi (2010), and Shen (2011) as applied to claim 1 above, and further in view of Liu (WO2017070633A2). This rejection is maintained.
The teachings of Dominguez-Monedero, Campbell, Hayashi, and Shen are discussed above and render obvious the limitations of claim 1. However, the Cas9 protein taught by Dominguez-Monedero does not have the sequence of any one of SEQ ID NOs:4-6.
Liu teaches Cas9 proteins of instant SEQ ID NOs:4-6.
Regarding claim 24, Liu teaches that wild-type Cas9 from Streptococcus pyogenes has the amino acid sequence of SEQ ID NO:2 (SEQ ID NO:2 of Liu is 100% identical to instant SEQ ID NO:4, see alignment below; paragraph [0096]).
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Further regarding claim 24, Liu teaches that D10A nickase Cas9 has the sequence of SEQ ID NO:301 (SEQ ID NO:301 of Liu is 100% identical to instant SEQ ID NO:5, see alignment below; paragraph [00101]).
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Regarding claim 25, Liu teaches that nuclease-inactive Cas9 D10A/H840A has the sequence of SEQ ID NO:262 (SEQ ID NO:262 of Liu is 100% identical to instant SEQ ID NO:6, see alignment below; paragraph [00122], claim 1).
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Liu teaches that nickase and nuclease-dead Cas9 variants, such as those described above, retain DNA binding functions and have value for genome engineering applications (paragraph [0008]). While Dominguez-Monedero and Campbell teach different Cas9 sequences (not instant SEQ ID NOs:4-6), it would have been obvious to a person of ordinary skill in the art at the time of filing that any known Cas9 sequence could be substituted for the Cas9 sequence of Dominguez-Monedero or Campbell, as changing the particular amino acid sequence of the Cas9 domain of the Cas9 fusion protein rendered obvious by the combined teachings of Dominguez-Monedero, Campbell, Shen, and Hayashi to a different wild-type, nickase, or nuclease-dead Cas9 sequence (such as those taught by Liu) would not prevent the addition of the myristoylation domain or the NLS domain, nor would prevent the packaging of the fusion protein in an extracellular vesicle.
Response to Arguments
Applicant's arguments filed 10/31/2025 have been fully considered but they are not persuasive.
Applicant first argues that the stated purpose of the myristoylation domain of Dominguez-Monedero is to target the Cas9-ERT2 fusion protein to the cell membrane in the absence of 4-hydroxytamoxifen, to provide a low basal level of Cas9 activity (page 7 of arguments). Applicant argues that the ERT2 or hPR domain of Dominguez-Monedero is required for the functioning of the Cas9 fusion protein of Dominguez-Monedero. However, that the myristoylation domain reduces but does not eliminate basal Cas9-hPR activity would indicate to an artisan that the myristoylation domain does not permanently bind the fusion protein to the cell membrane. In fact, Hayashi teaches that myristoylated proteins were known to be present in both the cell membrane and the cytoplasm, depending on further modifications (see Fig. 9, wherein phosphorylation is shown to dissociate myristoylated proteins from the cell membrane). An artisan would recognize that Dominguez-Monedero teaches multiple different and overlapping concepts: first, drug-inducible Cas9 fusion proteins, necessitating a drug-inducible domain such as ERT2 or hPR; second, Cas9 proteins with reduced basal activity, enabled by the fusion of the Cas9 to a myristoylation domain targeting the fusion protein to the cell membrane; and third, Cas9 proteins with an NLS domain for facilitated transport into the nucleus of a cell. While the stated novelty of Dominguez-Monedero is the drug-inducible Cas9 activity enabled by the drug-inducible domains taught, the separate utilities of the myristoylation domains and the NLS domains are described in Dominguez-Monedero and are apparent. An artisan would have recognized that elimination of the ERT2 or hPR domain would eliminate the drug-inducibility of the Cas9 function, but would not eliminate the membrane-targeting enabled by the myristoylation domain or the nuclear targeting enabled by the NLS domain.
Applicant then argues that “Hayashi teaches that differences in amino acid sequence at myristoylated domains altered their locations in cells, and that the target membranes may also be different”, and that therefore, it would not be reasonable to assume that all myristoylation domains were interchangeable (pages 7, 9). This argument is partially convincing. As the myristoylation domain of Dominguez-Monedero targets the fusion proteins to the cell membrane, it would be obvious to substitute another myristoylation domain which also targets proteins to the cell membrane. Given the benefits of introducing Cas9 as a ribonucleoprotein complex encapsulated in an extracellular vesicle compared to introducing Cas9 encoded on a plasmid (as taught by Campbell, described above), and given that Shen teaches a myristoylation domain which targets proteins to the cell membrane and to extracellular vesicle budding sites in the cell membrane, it would have been obvious to replace the myristoylation domain of Dominguez-Monedero, which is only taught to target proteins to the cell membrane, with a myristoylation domain which both targets proteins to the cell membrane and targets proteins to extracellular vesicles; the myristoylation domain of Shen would obviously fulfill the requirements for the myristoylation domain of Dominguez-Monedero (targeting the cell membrane) and the additional functional requirement of Campbell (targeting to extracellular vesicles). As such, while Applicant has successfully argued that not all myristoylation domains would be obvious to substitute, the Examiner sustains that the specific myristoylation domains of Dominguez-Monedero and Shen would be obvious to substitute. As a result of this argument, the rejections under 35 USC 103 of claims 10-11 and 23 have been withdrawn, as these myristoylation domains were not taught in the art to have the required function of targeting proteins to extracellular vesicles, and thus would not have been obvious to substitute for the myristoylation domain of Shen or Dominguez-Monedero.
Applicant argues that Dominguez-Monedero provides a strategy for reducing off-target Cas9 activity by the introduction of drug-inducible domains, and as such, the different strategy of Campbell for reducing off-target Cas9 activity (introduction to cells as an RNP encapsulated in an extracellular vesicle) would not be necessary, and it would not be obvious to replace one with the other. While the drug-inducible Cas9 fusion proteins of Dominguez-Monedero do provide a solution to off-target Cas9 activity, Campbell teaches other benefits provided by extracellular vesicle-mediated delivery of Cas9 which Dominguez-Monedero does not address. As discussed above, Campbell teaches that delivery of Cas9 as an RNP encapsulated in an extracellular vesicle provides cell type specificity and reduced toxicity compared to delivery to cells of Cas9 encoded in a plasmid. It thus would have been obvious to an artisan that modification of the proteins of Dominguez-Monedero to enable delivery by extracellular vesicle instead of by plasmid would provide additional benefits.
Regarding the rejections of claims 16-17 and 22, Applicant argues that the teachings of Chiper that NLS sequences target proteins to the nucleus of a cell is not universally true for all proteins, and that the “only conclusion that can be drawn from this is that the c-Myc NLS nuclear localization signal is useful for delivering GFP” (page 10). Applicant further argues that the GFP-NLS fusion proteins of Chiper require AuNPs for nuclear delivery. Chiper teaches that “[p]assive entrapment of large (>60 kDa) macromolecules into the nuclei occurs through mitosis, during which the nuclear compartment vanishes. Hence, for cell division independent nuclear import, critical in the case of proteins with exclusively nuclear activity, it is important to either respect a threshold limit (using <60 kDa proteins) or equip proteins with NLS to channel nuclear import using importin α/β machinery” (page 1701040). Chiper thus teaches that the benefit of an NLS domain is applicable to any protein, particularly proteins over 60kDa in size. Furthermore, Chiper teaches that the AuNPs are used for delivery of proteins into the cytoplasm, not into the nucleus (page 1701040 and Fig. 12). An artisan would recognize that the function of the AuNP and the NLS in Chiper were independent of each other, and the nuclear localization enabled by an NLS did not require a specific mechanism for delivery of a protein into the cytoplasm.
Regarding the rejections of claims 24-25, Applicant argues that it would not be obvious to substitute different Cas9 proteins, as each Cas9 has different activity and different PAM sequences and specificities; changing the Cas9 of Dominguez-Monedero for a different Cas9 would change the gene editing activity of the Cas9 fusion protein. Dominguez-Monedero, however, teaches that multiple CRISPR nucleases can be used in their invention (Abstract, page 2167, Dominguez-Monedero teaches using sp-Cas9 and hAsCpf1). It would have been obvious to an artisan that substituting a different Cas9 or Cpf1 sequence would not eliminate the RNA-guided DNA-binding function required of the CRISPR nuclease domain by Dominguez-Monedero. In fact, substitution of different Cas9 domains with different activities and different PAM sequences would enable targeting of different sequences. Furthermore, as the teachings of Dominguez-Monedero which obviate the claimed invention do not require specific gRNA target sequences or Cas9 activity levels, and instead relate to the addition of domains providing cell membrane- and nucleus-targeting functionalities, changes in Cas9 activity levels or PAM sequences would not fundamentally alter the relevant activities and functions of the Cas9 fusion proteins taught by or rendered obvious by Dominguez-Monedero (cell membrane targeting, nucleus targeting, RNA-guided DNA binding and/or cleavage).
As such, the above rejections of the pending composition claims are held to be proper and are maintained.
Allowable Subject Matter
Claims 10-11 and 23 are objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims.
The following is a statement of reasons for the indication of allowable subject matter: Applicant has argued that different myristoylation domain sequences provide different localization and function, and that as a result, myristoylation domains cannot be assumed to be interchangeable if specific localization or function is required. The prior art does not teach that myristoylation domains comprising SEQ ID NOs:340-341 or encoded by SEQ ID NO:402 target proteins to the cell membrane and to extracellular vesicles. As such, it would not have been obvious to an artisan to use myristoylation domains comprising SEQ ID NOs:340-341 or encoded by SEQ ID NO:402 in the Cas9 fusion proteins rendered obvious by the combination of Dominguez-Monedero, Campbell, Hayashi, and Shen, as described in the rejection of claim 1 under 35 USC 103 above.
Conclusion
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/AFRICA M MCLEOD/ Examiner, Art Unit 1635
/KIMBERLY CHONG/ Primary Examiner, Art Unit 1636