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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 04/02/2026 has been entered.
Application Status
This action is written in response to applicant’s correspondence received 04/02/2026. Claims 1-2, 8-10, 12, and 14-25 are currently pending. No claims are withdrawn from prosecution as being drawn to non-elected subject matter. Accordingly, claims 1-2, 8-10, 12, and 14-25 are examined herein.
Any rejection or objection not reiterated herein has been overcome by amendment. Applicant' s amendments and arguments have been thoroughly reviewed, but are not persuasive to place the claims in condition for allowance for the reasons that follow.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
Determining the scope and contents of the prior art.
Ascertaining the differences between the prior art and the claims at issue.
Resolving the level of ordinary skill in the pertinent art.
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, 9-10, 12, 14, 16-20 and 22 are rejected under 35 U.S.C. 103 as being unpatentable over Feng et al. (Lytic Induction Therapy for Epstein-Barr Virus-Positive B-Cell Lymphomas. J Virol. 2004 Feb;78(4):1893–1902.) in view of WIPO Publication 2017/058795 to Quake (applicant’s submission, hereinafter ‘Quake’).
Regarding claims 1-2, 9 and 14:
Feng et al. teach a method for treating EBV-positive tumors which comprises activating lytic EBV by introducing into the cell a transcriptional activator (i.e., gemcitabine and doxorubicin) that is capable of activating transcription of lytic EBV genes (the two viral immediate-early transactivator genes, BZLF1 and/or BRLF1)(§Abstract).
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Please note that Feng et al.’s method comprised activating transcription from the promoters, i.e., transcriptional regulatory sequences (see above).
Feng et al. further note that the ability of various chemotherapy drugs to induce lytic EBV induction is cell type dependent:
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Feng et al. further provide a teaching, suggestion or motivation to use gene therapy/delivery methods to induce lytic EBV infection by expressing the BZLF1 or BRLF1 genes (p. 1901):
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Feng et al. do not teach that the method of activating EBV uses a programmable DNA binding protein system (dCas9) that is linked to a transcriptional activator.
Quake teaches methods of upregulating viral transcription using a Cas9 system targeting a viral promoter:
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Quake further teaches that EBV promoters may be targeted with said method:
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Quake further teaches activation of viral transcription by fusing a transcriptional activator to dCas9 (p. 10/30) (relevant to claims 1 and 9-10):
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Quake further teaches that the system may be delivered via AAVs, which allow for preferential infection of certain tissues and would have addressed the issue of cell type dependence noted by Feng et al. (p. 15/30):
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It would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of activating lytic EBV in tumor cells to treat EBV-driven lymphoproliferative disease via administration of gemcitabine and GCV , as taught by Feng et al., to induce the lytic form of EBV via introduction of dCas9 fused to a transcriptional activator, as taught by Quake. The skilled artisan would have been motivated by Feng et al. to induce lytic EBV, via transcriptional activation of viral lytic genes, as a way to treat EBV-associated tumors, but to do so via gene therapy rather than administration of chemotherapeutic drugs. As Feng et al. note, the chemotherapy-based method has the limitation that it is cell type dependent. Quake, instead, provides an alternative method for activating lytic EBV by using a dCas9/transcriptional activator system which may be delivered via AAV, which would have permitted preferential infection of target tissues. One having ordinary skill would have recognized that Quake’s method followed Feng et al.’s suggestion of a gene therapy approach while overcoming the limitations of Feng et al.’s chemotherapy-based approach.
Regarding claim 12, Quake teaches wherein the programmable DNA binding protein system comprises a transcription activator-like effector (TALE) or zinc finger nuclease (ZFN) (p 9/30):
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Regarding claim 14, Quake and Feng et al. both teach wherein the transcriptional activator binds to the transcriptional regulatory sequence (promoter) (see above).
Regarding claims 16-18, Feng et al. teaches that the cell is a mammalian and/or cancer cell (Abstract; lymphoblastoid cells), and teaches the introduction of an antiviral prodrug, ganciclovir (GCV) into the cell (Abstract and p. 1893):
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Regarding claim 19, it is noted that the claim recites the same limitations present in claims 1-4 and 14 with regard to the method steps, genes targeted, and gene regions targeted. Claim 19 differs from the previous claims in that it further recites intended outcomes of the same method steps, i.e., at least 2-fold higher expression of BZLF1 and BRLF1 when both genes are targeted simultaneously compared to a method in which either gene is targeted alone. However, the limitation is not considered to have patentable weight over the prior art method which renders the method steps obvious, because specification does not appear to show at least a 2-fold increase in expression of BZLF1 and BRLF1 when transcription of both genes is activated.
Figure 11 (depicting the results discussed in Example 6 on p. 36) shows a gel comparing relative protein expression after administration of either sgRNA (BZLF1 or BRLF1) alone to both at once. However, the amounts are not quantified, and the strength of the bands does not clearly show a 2-fold increase. For example, the band intensity for BZLF1 is stronger in the BZLF1 sgRNA (A5) only lane (lanes 2 and 6) than in the BZLF1 + BRLF1 lanes (4 and 8), which would indicate that protein expression was higher for the single BZLF1 sgRNA than it was for the BZLF1 and BRLF1 sgRNAs combined. Example 13 on p. 41 discusses the synergistic activation of both genes. Applicant defines “synergistic EBV lytic activation” as, “activation of multiple EBV lytic genes.” (p. 15 ln 5). This is interpreted to encompass any non-zero amount of activation. The specification states that synergistic activation “may” be 2-fold to 100-fold relative to non-synergistic activation, but the use of the term “may” is exemplary, open-ended, and does not disavow the full scope of the term (MPEP 2111.01(IV). Example 13 does not describe the level of activation, stating only that the proteins “were detectable” (p. 41 ln 12), that synergistic protein production occurred, and that the co-expression of BZLF1sgRNA3 and BRLF1 sgRNA3 increased the number of cells expressing caspase-3, more cells were accumulated in sub G1 phase, and the cell viability was decreased compared with DOX treated dCas9-Tet on - p65HSF1-BZLF1 sgRNA3 SNU-719, C666-1, and C17 cells (p. 41 ln 14-17). Absent support in the specification which shows, “wherein expression of genes regulated by EBV BZLF1 and EBV BRLF1 is at least 2-fold higher than expression of the same genes resulting from introduction of a programmable DNA binding protein system that targets only EBV BZLF1 or only EBV BRLF1”, the limitation is not given patentable weight for the purposes of comparison to the prior art.
Regarding claims 20 and 22, Feng et al. teaches administering the system for inducing lytic EBV to the subject (and thus also teaches cells comprising the system), wherein the subject has cancer associated with EBV infection (§GCV enhances the therapeutic effect of gemcitabine and doxorubicin in EBV-positive lymphoproliferative disease in SCID mice).
Claim 8 is rejected under 35 U.S.C. 103 as being unpatentable over Feng et al. and Quake as applied to claims 1-2, 9-10, 12, 14, 16-20 and 22, further in view of Konermann.(Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature volume 517, pages583–588 (2015).; of record, applicant’s submission).
Feng et al. and Quake render obvious the method of claim 1, from which instantly rejected claim 8 depends, as described above
Feng et al. and Quake do not teach wherein the transcriptional activator comprises a heat shock factor transactivation domain.
Konerman teaches a transcriptional activation system comprising dCas9 and a transcriptional activator which comprises HSF1 and is linked to a component of the complex. Please see Figure 1b, which shows a dCas9-VP64 fusion protein complexed with sgRNA, which is linked to a MS2-p65-HSF1 fusion protein. In the figure caption, Konerman notes, “Addition of the HSF1 transactivation domain to MS2-p65 further increases the efficiency of the transcription activation”.
It would have been prima facie obvious to a person having ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of claim 1 by substituting the generic transcriptional activator with a transcriptional activator comprising HSF1. The skilled artisan would have been motivated to do so, and would have had a reasonable expectation of success, based on Konerman’s teachings that addition of the HSF1 transactivation domain enhanced the efficiency of the transcription activation.
Claims 23-25 are rejected under 35 U.S.C. 103 as being unpatentable over Feng et al. and Quake as applied to claims 1-2, 9-10, 12, 14, 16-20 and 22, further in view of Cheng et al. (Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cell Res. 2016 Feb;26(2):254-7.; applicant’s submission).
Feng et al. and Quake render obvious the method of claim 1, from which instantly rejected claims 23-25 depend, as described above.
Feng et al. and Quake do not teach wherein the transcriptional activator comprises P65HSF1 (relevant to claim 25), or wherein the Cas9 system further comprises a PUF domain binding sequence (PBS) linked to the guide RNA and a PUF domain that binds to the PBS of the gRNA, and the PUF domain is linked to the transcriptional activator (claims 23-24).
Cheng et al. teach the Casilio system, a CRISPR-Cas9-Pumilio hybrid having the recited components:
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Cheng et al. further teach that they were able to achieve “robust activation…in the corresponding Casilio experiments” (p. 256 first ¶; Figure 1C).
It would have been prima facie obvious to a person having ordinary skill in the art before the effective filing date of the claimed invention to have modified the method for activating a lytic Epstein-Barr virus gene comprising introducing into the cell a programmable DNA binding system linked to a transcriptional activator, as taught by Feng et al. and Quake, by using the transcriptional activator comprising P65HSF1/a PUF domain binding sequence (PBS) linked to the guide RNA and a PUF domain that binds to the PBS of the gRNA, and the PUF domain is linked to the transcriptional activator, as taught by Cheng et al., in place of the generic transcriptional activator taught by Feng et al./Quake. The skilled artisan would have had a reasonable expectation of success based on Cheng et al.’s successful application of the Casilio system to achieve transcriptional activation of target genes.
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Feng et al. and Quake as applied to claims 1-2, 9-10, 12, 14, 16-20 and 22, further in view of Guo et al. (An inducible CRISPR-ON system for controllable gene activation in human pluripotent stem cells. Protein Cell 2017, 8(5):379–393.).
Feng et al. and Quake render obvious the method of claim 1, from which instantly rejected claim 15 depends, as described above. Specifically, Feng et al. in view of Quake teach a system with a dCas9 protein fused to a transcriptional activator (see above).
Feng et al. and Quake do not teach wherein expression of the DNA binding protein and/or transcriptional activator is inducible.
Guo et al. teach an inducible CRISPR-ON system for controllable gene activation (Title), in which a dCas9 protein is fused to a transcriptional activator and placed under the control of an inducible promoter. The system comprises a doxycycline inducible dCas9-VP64-p65-Rta transcription activator (§Abstract). The DCas9-VPR was constructed by fusing the nuclease deficient Cas9 (dCas9) with transcription activator VP64, p65 and Rta in tandem (p. 390 §Plasmid construction). For inducible expression, dCas9-VPR was placed behind a TRE promoter (p. 391 §Plasmid construction first ¶). Guo et al. note that the system can be a valuable system to control gene expression from endogenous loci (p. 380 second ¶).
It would have been prima facie obvious to a person having ordinary skill in the art before the effective filing date of the claimed invention to have modified the method as taught by Feng et al. and Quake by making the expression of the DNA binding protein and/or transcriptional activator inducible. The skilled artisan would have been motivated to do so by Guo et al.’s teachings, which demonstrated that a system placing the dCas9/transcription activator fusion protein under the control of an inducible promoter was effective and could be a valuable system to control gene expression from endogenous loci, such as latent viral genomes in EBV-infected cells.
Claim 21 is rejected under 35 U.S.C. 103 as being unpatentable over Feng et al. and Quake as applied to claims 1-2, 9-10, 12, 14, 16-20 and 22, further in view of Ahern (Biochemical, Reagents Kits Offer Scientists Good Return On Investment. The Scientist, Vol:9, #15, pg 20, July 24, 1995).
Feng et al. and Quake render obvious the method of claim 1, from which instantly rejected claim 21 depends, as described above.
Feng et al. and Quake do not teach a kit comprising the system of the method of claim 1.
Ahern teaches that premade reagents and kits are convenient and save time for researchers. Rather than browsing through catalogs and buying individual chemicals from one or several suppliers, investigators can instead purchase a kit that supplies all of the necessary reagents for a particular research application and even provides them with detailed instructions (p. 5/10 last ¶).
It would have been prima facie obvious to a person having ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of claim 1, as taught by Feng et al. and Quake, by including the necessary components in a kit so that the method could be carried out with greater ease and in less time, as taught by Ahern.
Response to Arguments
Applicant's arguments filed 04/02/2026 have been fully considered but they are not persuasive because a) there is no nexus between the absence of unexpected results and the claimed invention, and b) the evidence of unexpectedly superior results is not commensurate in scope with the claimed invention. A more detailed explanation follows.
Applicant argues, “Example 7 shows that targeting a third EBV lytic gene, BGLF4, did not achieve the same level of success. Viewed together, these results indicate that, regardless of whatever relevant teaching one might derive from Feng and Quake, not all EBV lytic genes that could be targeted would, upon being activated, in fact turn out to successfully induce lysis of the EBV+ host cells.”.
“The absence of a property which a claimed invention would have been expected to possess based on the teachings of the prior art is evidence of unobviousness.” (MPEP 716.02(a)). In the instant case, Applicant’s argument is understood as indicating that the absence of successful reactivation of EBV by targeting BGLF4 was unexpected based on Feng’s teachings. This argument is acknowledged. Respectfully, however, it is not persuasive because there is insufficient nexus between the claimed invention and the evidence of unexpected results.
MPEP 716.01(b) states, “If the evidence is to be given substantial weight in the determination of obviousness or nonobviousness, evidence of secondary considerations must be relevant to the subject matter as claimed, and therefore the examiner must determine whether there is a nexus between the merits of the claimed invention and the evidence of secondary considerations.”. Further, “"Where the offered secondary consideration actually results from something other than what is both claimed and novel in the claim, there is no nexus to the merits of the claimed invention.”. In the instant case, the claimed invention does not encompass targeting of BGLF4, so the offered secondary consideration results from something other than what is both claimed and novel in the claim.
Applicant further argues, “Applicant contends that the claimed method of using EBV lytic gene BZLFI or BRLFI to induce the lysis of EBV+ cells must be recognized as non-obvious because of the unexpected beneficial outcome and technical advantages it is able to achieve.” Applicant provides, as evidence, the Wu et al. publication (attached to the Remarks as Exhibit 1), noting, “poorer efficiency of various chemical inducers in inducing EBV lytic proteins is shown (suppl. Figure 1), whereas the CRISPR- and TALE-systems (the claimed method of this application) are able to induce the lytic reaction in all EBV-positive cancer cell lines (Figure 1).”. Respectfully, this argument is not persuasive because the evidence of unexpectedly superior results is not commensurate in scope with the claimed invention (MPEP 716.02(d)).
Claim 1 is drawn to a method of activating a lytic EBV, comprising introducing into a cell infected with EBV a generic programmable DNA binding protein system that targets a transcriptional regulatory sequence of a lytic EBV gene BZLF1 or BRLF1 (e.g., a promoter or enhancer), and a transcriptional activator that is linked to a component of the programmable DNA binding protein system and is capable of activating transcription of the lytic EBV gene. The claim is generically broad as to the components of the system and the activator, encompassing systems including but not limited to any Cas-based system in which a catalytically dead Cas9 is fused to any transcriptional activator (e.g., the dCas9 Casilio system described on p. 12 of Wu et al.), as well as TALE systems (also described in Wu et al.). However, the claim does not recite any specific type of programmable DNA binding protein, any specific type of transcriptional activator, or any specific sequence required to achieve the claimed outcome, such as a guide RNA sequence or a TALE recognition site sequence or the coding sequence of a TALE protein capable of recognizing said recognition site sequence.
In contrast to the breadth of the claims, the evidence in the instant specification and in Wu et al. is relatively narrow.
Beginning with the specification:
The specification discloses that lytic reactivation was tested by transfecting, “2 micrograms ( g) of control gRNA or individual BZLF1 gRNA (A3) (SEQ ID NO: 8), gRNA (A4) (SEQ ID NO: 9), gRNA (AS) (SEQ ID NO: 10), or gRNA (A6) (SEQ ID NO: 11) gRNAs,” into, “SNU-719 and C666-1 cells stably expressing HA-dCas9-EGFP and 3XFLAG-PUFa- p65HSF1” (p. 31 ln). In other words, a specific dCas9 Casilio system, as described in Wu et al., was used with specific gRNAs.
The results were as follows: “BZLF1 gRNA (A5) and BZLF1 gRNA (A6) reactivate EBV expression at detectable levels in both C666-1 and SNU-719 cells”; “BZLF1 gRNA (A3) and BZLF1 gRNA (A4) also reactive EBV expression at detectable levels in SNU-719 cells”, and “BZLF1 gRNA (A5) activated BZLF1 (Zta) protein expression in both C666-1 and SNU-719” (p. 31 ln 15-30). As seen in FIG. 7A, sgRNAs A3 and A4 showed no detectable Zta protein expression in C666-1 cells. However, as seen in FIG. 7B, there was at least minimal Zta protein expression detected after administration of any of A3-A6 in SNU-719 cells. Overall, it appears that BZLF1 gRNA A5 and A6 showed detectable EBV activation in at least two different cell lines, but the other gRNAs did not. This evidences that not all gRNAs will predictably, successfully reactivate EBV in all cell types. Because the claims generically encompass reactivation of EBV with any gRNA in any cell, or indeed any transcriptional activation system, the unexpected results in the form of successful activation of BZLF1 is not commensurate in scope with the claims. Similarly, the specification discloses reactivation of EBV with a Casilio system and specific guide RNAs (2) and (3) targeting BRLF1, but not with the other BRLF1 gRNAs (1, 4, 5, or 6) tested. The sequences of gRNAs BZLF1 A5 and A6 are SEQ ID NOs: 10 and 11, respectively (Table 1, p. 47). The sequences of gRNA BRLF1 (2) and (3) are SEQ ID NOs: 13 and 14, respectively (Id.).
The claims also encompass TALE nucleases linked to transcriptional activators. In Example 8, the specification discloses TALEs targeting BZLF1, but not BRFL1, and only discloses successful reactivation using BZLF1 TALE (3) (p. 37 ln 10-30). In Example 15, the specification discloses testing “the utility of TALE transactivator for activating BZLF1 expression” and tests four BZLF1 TALE activators. However, the specification notes that, “TALE (3) gave the highest activity in both cells, while TALE (2) was only able to activate BZLF1 in C666-1 cells instead of and [sic] SNU-719 cells” (p. 42 ln 10-15). Based on FIGs. 22A and 22B, TALE BZLF1 (3) was able to induce robust BZLF1 expression in both cell lines, followed by TALE BZLF1 (4), and TALE BAZLF1 (2) was indeed only able to induce BZLF1 expression in SNU-719 cells. Regarding BRLF1, in Example 16, a, “TALE transactivator targeting BRLF1 promoter was constructed and transiently transfected into C666-1 cells, resulting in the activation of BRLF1 as well as BZLF1 (FIG. 23A). The recognition sites for TALE BZLF1 (3) and (4) are represented by SEQ ID NOs: 27 and 28, respectively (Table 2, p. 48). The amino acid sequence of BZLF1 TALE (3) is SEQ ID NO: 6 (p. 45 ln 5-30). The recognition site for TALE BRLF1 is SEQ ID NO: 29 (Table 2, p. 48) and the amino acid sequence of TALE BRLF1 is SEQ ID NO: 32 (p. 46 ln 5-30).
Regarding Wu et al.:
Wu et al. also discloses EBV lytic activation using a Casilio system and gRNAs. However, they show the same unpredictability as the specification: for example, sgRNA1 (equivalent to the instantly closed gRNA (A3), SEQ ID NO: 8 – see Supplementary Table 1 of Wu et al. and Table 1 of the specification) showed no detectable Zta in C666-1 lines. Wu et al. does not disclose gRNAs targeting of BRLF1 at all. Similarly, Wu et al. discloses successful EBV lytic activation using a TALE plasmids targeting BZLF1: TZ1, TZ2, TZ3 and TZ4 (Fig. 2, especially 2a and 2b). The results were variable depending on cell line, with TZ1 showing low to no activation and TZ3 showing the highest. According to Supplementary Table 1 of Wu et al., the TALE recognition site sequences were essentially the same as those disclosed in Table 2 of the specification, except that Wu’s sequences have an additional ‘T’ at the 5’ end.
Further, it is relevant to note that, according to Wang et al., the effects of targeting EBV regulatory sequences can differ widely and in unpredictable ways, noting, “CRISPRi targeting of an enhancer can give up to 50% reduction of target gene transcript levels (Figure 3B; see Table 3 for sequences of validation primers). However, the effects can vary widely between different genomic loci for several reasons. First, CRISPRa and CRISPRi sgRNAs are targeted towards non-protein-coding regions which have not been under the same evolutionary pressures as protein-coding gene bodies. Hence, CRISPRa and CRISPRi sgRNAs are expected to be more promiscuous in their binding of regulatory elements on DNA, yielding substantial variation in gene expression modulation. Second, nucleosome positioning may influence Cas9 activity; if the PAM site is obscured within the nucleosome core, Cas9-sgRNA RNPs can become restricted in their ability to recognize cognate sequences.” (Wang et al. Modulating Gene Expression in Epstein-Barr Virus (EBV)-Positive B Cell Lines with CRISPRa and CRISPRi. Curr Protoc Mol Biol. 2018 Jan 16;121:31.13.1–31.13.18. ). While Wang et al. focused on transcriptional repression, not activation, they make clear that the unpredictability is due to the structure of the target sites and sgRNAs and the interactions between them. This further evidences that not all gRNAs or TALEs targeting EBV regulatory elements will predictably yield the same results.
In summary, while the instant specification provides evidence that specific programmable DNA binding proteins linked to specific transcriptional activators and using specific gRNA or TALE recognition sites/TALEs, and while Wu et al. provides additional support that certain of the specific sgRNAs and TALEs were more effective than chemical inducers, the evidence is limited to specific constructs and is not commensurate in scope with the broader claims.
Subject matter free of the prior art
The prior art does not teach or suggest:
A method for activating a lytic Epstein-Barr virus (EBV) gene, comprising introducing into a cell infected with EBV:
i) a programmable DNA binding protein system that targets a transcriptional regulatory sequence of a lytic EBV gene BZLF1 or BRLF1;
ii) and a transcriptional activator that is linked to a component of the programmable DNA binding protein system and is capable of activating transcription of the lytic EBV gene;
wherein:
a) the programmable DNA binding protein system is a Cas9-based system, comprising a ribonucleoprotein complex comprising a catalytically-inactive Cas9 nuclease bound to a guide RNA (gRNA) targeting BZLF1 or BRLF1, wherein the gRNA targeting BZLF1 comprises the sequence of SEQ ID NO: 10 or SEQ ID NO: 11, the gRNA targeting BRLF1 comprises the sequence of SEQ ID NO: 13 or SEQ ID NO: 14; and the gRNA targeting BZLF1 or BRLF1 is linked to a Pumilio-FBF (PUF) domain binding sequence (PBS), and the PBS is bound to a PUF domain that is linked to a p65HSF1 transactivation domain; or,
b) the programmable DNA binding protein system is a TALE-based system targeting BZLF1 or BRLF1, comprising a transcription activator-like effector (TALE) linked to the transcriptional activator, wherein the TALE-based system targeting BZLF1 comprises the amino acid sequence of SEQ ID NO: 6 and the TALE-based system targeting BRLF1 comprises the amino acid sequence of SEQ ID NO: 32.
As discussed in the rejections of the claims under 35 U.S.C. 103, the closest art to the claimed invention is represented by Feng in view of Quake. However, as discussed in the response to arguments above, the specification provides objective evidence of nonobviousness in the form of both unexpectedly superior results and by evidencing unpredictability in the effects of different gRNAs and TALEs on EBV reactivation. Based on this evidence, systems which use the specific gRNAs and TALEs noted above to reactivate lytic EBV are considered nonobvious over the prior art.
Conclusion
No claim is allowed at this time.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to AMANDA M ZAHORIK whose telephone number is (703)756-1433. The examiner can normally be reached M-F 8:00-16:00 EST.
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/AMANDA M ZAHORIK/ Examiner, Art Unit 1636
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