DETAILED ACTION
Claims 1-9 and 19-21 are pending.
Status of Claims
Claims 1-9 and 12-21 are pending. Claims 1 and 7 have been amended. Claims 1-9 and 12-21 are under examination.
Claim Rejections - 35 USC § 103-New: Necessitated by Amendments.
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.
1.Claim 1-5 and 19-20 are rejected under 35 U.S.C. 103 as being unpatentable over Fuenzalida et al., (WO 2017129811 A1) (IDS filed on 10/31/2022), in view of Pedersen (WO 2015052231 A2) (IDS filed on 10/31/2022), and in view of Doench, John G et al. “Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation.” Nature biotechnology vol. 32,12 (2014): 1262-7. doi:10.1038/nbt.3026 (List of references cited on 08/27/2024).
Fuenzalida teaches a method comprising a. administering Cas-9 to a population of cells, wherein Cas-9 is administered using CRISPR technology, where gRNA against a marker gene, gRNA against a target sequence are administered using CRISPR technology to the same
population of cells prior to a negative selection step (see [0063] “In a further preferred
embodiment the invention provides an in vitro method for enriching eukaryotic cells which are
modified by homologous recombination, comprising (a) subjecting a population of cells
transformed with a nucleic acid molecule or a composition of the invention to means for
selecting for said marker indicating heterologous recombination and separate transformed cells
expressing said selection marker indicating heterologous recombination in said eukaryotic cell;
and (b) subjecting the non- separated cells to means for selecting for said marker indicating
homologous recombination in order to enrich transformed cells comprising said homologous
recombination." See [0036] “The nucleotide sequence encoding a selection marker indicating
homologous recombination may be removed by excision or recombination or cleavage after
depositing a modification into the genome. The removal of the nucleotide sequence encoding a
selection marker indicating homologous recombination may be performed by the use of a
recombinase, transposase, RNA guided nuclease or nuclease.”, see [0048] “The term "CRISPR
Cas" as used herein relates to the Clustered Regularly Interspaced Short Palindromic Repeats
(CRISPR) Type II system which is a bacterial immune system that has been modified for
genome engineering. CRISPR consists of two components: a "guide" RNA (gRNA) and a non-specific CRISPR-associated endonuclease (Cas9). The gRNA is a short synthetic RNA
composed of a "scaffold" sequence necessary for Cas9-binding and a user-defined ~20
nucleotide "spacer" or "targeting" sequence which defines the genomic target to be modified.
Thus, one can change the genomic target of Cas9 by simply changing the targeting sequence
present in the gRNA. CRISPR/Cas can be used for gene engineering by co-expressing a gRNA
specific to the sequence to be targeted and the endonuclease Cas9.”, see [0048] teaching the
use of CRISPR Cas-9 and gRNA being administered together, thus teaching them being applied
to the sample population),
b. performing FACS-based negative selection on the population of cells to establish an
enriched cell population of negatively selected cells (see [0036] “Cells that have undergone
removal of the nucleotide sequence encoding a selection marker indicating homologous
recombination may further be enriched by the selection of cells that lost the fluorescence signal
by FACS or fluorescence microscopy.”); wherein the negatively selected cells do not comprise a
marker encoded by the marker gene and do comprise a mutation in the target sequence (see
[0064] “In the second selection step (b) non-separated cells (cells which are not positive for the
selection marker indicating heterologous recombination) are selected for said marker indicating
homologous recombination indicating a site-specific integration of the nucleic acid molecule or
the composition of the invention. Said second selection step separates cells which do not
comprise the selection marker, indicating that said cells were not successfully transformed.”).
Fuenzalida teaches encoding a selection marker (see abstract “The present invention
relates to a nucleic acid molecule at least one nucleotide sequence encoding a selection marker
indicating homologous recombination in a eukaryotic cell and at least one nucleotide sequence
encoding a selection marker indicating heterologous recombination in said eukaryotic cell.”)
(instant claim 1). Fuenzalida teaches wherein the CRISPR technology knocks out the marker gene (see [0036] “Cells that have undergone removal of the nucleotide sequence encoding a selection marker indicating homologous recombination may further be enriched by the selection of cells that lost the fluorescence signal by FACS or fluorescence microscopy.”, see [0062] “one nucleic acid molecule comprises a selection marker gene”) and mutates the target sequence ([0064] “non-separated cells (cells which are not positive for the selection marker indicating
heterologous recombination) are selected for said marker indicating homologous recombination
indicating a site-specific integration of the nucleic acid molecule or the composition of the
invention.”, [0039] “Such a mismatch in the homology arms may be employed in order to
introduce mutations in the nucleic acid sequence of interest.”) (instant claim 2).
Fuenzalida does not teach towards the use of sgRNA and the maker gene encoding a
cell surface protein B2M and the cell surface protein not being essential for cell survival. Fuenzalida does not teach the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence being expressed on different constructs, nor FACs based negative selection, nor do they teach the antibody being an anti-MHC I antibody.
Pedersen teaches multiplex editing systems (see abstract) and the use of sgRNA against a marker gene (see pg. 98 lines 1-5 “The targeted integration system is based
on three DNA parts: (a) a vector expressing Cas9 which is 2A-linked to a fluorescent marker (for
example GFP); (b) donor DNA with homology arms towards the integration site containing an
expression cassette inside the donor arms and a fluorescent marker gene (for example
mcherry) outside the homology arms; and (c) a sgRNA targeting the selected integration site
(Figure 18).”). Pedersen teaches the use of CRISPR Cas-9 (pg. 21 lines 9-11 “In preferred
embodiments, the endonuclease or variant thereof is Cas9 or a variant thereof, and the
targeting means are gRNAs that enable precise targeting of Cas9 to the TES.”, abstract “Also
provided herein are a method for editing nucleic acids and a cell comprising a stably integrated
endonuclease.”, pg. 21 lines 9-12 “In preferred embodiments, the endonuclease or variant
thereof is Cas9 or a variant thereof, and the targeting means are gRNAs that enable precise
targeting of Cas9 to the TES. Cas9 is a CRISPR-associated nuclease originally discovered in
Streptococcus pyogenes.”) (instant claim 1).
Pedersen teaches wherein the marker gene encodes b-2 microglobulin (B2M) (see pg.
41 lines 32-35 “For experiment part 4 we are constructing several CRESC's each with an FP as SM and a "housekeeping gene" (e.g. GADPH, B2M, etc.) because they are easy/cheap to
detect/quantify with RT-qPCR and not likely to cause disturbances in the cells.”) (Instant claim
5). Pedersen teaches a marker gene that encodes b-2 microglobulin (B2M) (see pg. 41 lines 32-35). B2M is a major histocompatibility complex (MHC) class I molecule. MHC class I molecules are found on the cells surface. B2M is inherently a cell surface protein Thus, Pedersen teaches wherein the marker gene encodes a cell surface protein (see pg. 41 lines 32-35). It would have been obvious that the B2M is not essential for cell survival because there are known cells, such as red blood cells, that exist without B2M (instant claims 3-4).
Doench teaches wherein the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed from different constructs (see pg. 1262 “These sgRNAs were cloned as a pool into a lentiviral vector that simultaneously delivers CRISPR-associated protein (Cas)9, confers puromycin resistance, and expresses a sgRNA, as previously described. A second pool targeting the coding sequence of three human cell surface markers and also including negative controls was separately cloned into a lentiviral vector that expresses only the sgRNA (Fig. 1a and Supplementary Table 2).”, see page 1262 "We designed sgRNAs that targeted a panel of mouse genes in all exons and all flanking intronic sequences at all 20 nucleotide (nt) target sites that preceded the NGG PAM required by S. pyogenes Cas9 and added a large number of negative control sgRNAs (Fig. 1a and Supplementary Table 1)”. The marker gene is human, whereas the target gene is mouse, see pages 1263-1264) (instant claim 1). Doench teaches the sgRNA against a marker gene targets an endogenous marker gene (see figure 2, see page 1264 “We next examined the consistency across cell lines of the activity of sgRNAs targeting CD13 or CD33. We observed strongly corre-lated sgRNA activity in four pairwise cross-cell-line comparisons, suggesting that relative levels of sgRNA activity can generalize across cellular contexts”, see page 1268 “Antibodies used in this study included: BD Pharmingen 555450 CD33-PE; BD Pharmingen 555394 CD13-PE; BD Pharmingen 562371 CD15-PE.”, see abstract “We created a pool of sgRNAs, tiling across all possible target sites of a panel of six endogenous mouse and three endogenous human genes and quantitatively assessed their ability to produce null alleles of their target gene by antibody staining and flow cytometry.”) (instant claim 1).
Doench teaches wherein FACS-based negative selection comprises administering an antibody capable of binding to the marker (see fig. 1, see pg. 1262 “Our strategy was to target cell surface markers in a large cell population, delivering one sgRNA per cell, and then isolating complete (biallelic) knockout cells by fluorescence-activated cell sorting (FACS), thereby separating the most active sgRNAs. We designed sgRNAs that targeted a panel of mouse genes in all exons and all flanking intronic sequences at all 20 nucleotide (nt) target sites that preceded the NGG PAM required by S. pyogenes Cas9 and added a large number of negative control sgRNAs (Fig. 1a and Supplementary Table 1) …We transduced EL4 cells, a mouse thymic cell line, with the mouse sgRNA pool. Nine days after transduction, we stained cells for each of nine cell surface markers and analyzed them by FACS. Endogenous Thy1, H2-K, Cd45, Cd43, Cd28 and Cd5 exhibited good resolution of marker-negative cell populations (Fig. 1b), whereas Cd2, Cd3e and Cd53 were poorly expressed and excluded from subsequent analyses (Supplementary Fig. 1).”, Online Methods “FACS. Human and mouse cell surface markers were selected the basis of homogeneity of expression as assessed by antibody staining profiles. Only cell lines which showed expression of a particular cell surface marker in >98% cells were chosen for analysis. EL4 cells were independently stained and sorted on a FACS Aria flow cytometer 8 d after transduction. Antibodies used in this study included: eBioscience 17-5958-80 Anti-Mouse MHC Class I (H- 2Kb) APC”). MOLM13, NB4 and TF1 cells were stained and sorted on a BD-FACS Aria II 8 d after transduction with the human sgRNA library. Antibodies used in this study included: BD Pharmingen 555450 CD33-PE; BD Pharmingen 555394 CD13-PE; BD Pharmingen 562371 CD15-PE.”) (instant claim 19). Doench teaches wherein the antibody is an anti-MHC I antibody (see Online Methods “Antibodies used in this study included: eBioscience 17-5958-80 Anti- Mouse MHC Class I (H-2Kb) APC”).” (instant claim 20).
It would have been obvious to one of ordinary skill in the art at the time the application
was filed to modify Fuenzalida’s methods of selecting transformed cells with the methods of using sgRNA for multiplexing taught by Pedersen, and with Doench’s teachings of using Cas-9 and sgRNA’s from different constructs. Fuenzalida teaches that when CRISPR-Cas9 is used a point mutation in the protospacer adjacent motif (PAM) sequence is introduced (see [0082], and that the PAM sequence is absolutely necessary for target binding and the exact sequence is dependent upon the species of Cas9 (see [0048]). Pedersen that their methods would be used to allow multiple insertions of genes of interest, i.e., multiplex editing of nucleic acids, in particular of genomes, by allowing repeated use of advantageous target locations”). Doench teaches that CRISPR-Cas9 programmed with a single guide RNA (sgRNA) to generate site-specific DNA breaks (abstract). Doench further teaches that using a sequence that correlates to on-target activity of the Cas9: sgRNA complex enables more effective applications of CRISPR technology when editing the genome and probe gene function (pg. 1266). It would have been obvious to one of ordinary skill in the art at the time of the application to use constructs that express Cas-9, sgRNA against a marker gene and target sequence. The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
2.Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Fuenzalida,
Pedersen, and Doench as applied to claims 1-5 and 19-20 above, and in further view of Oji et al. “CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice.” Scientific reports vol. 6 31666. 17 Aug. 2016, doi:10.1038/srep31666 (2016) (List of references cited on 08/27/2024).
The teachings of Fuenzalida, Pedersen, and Doench as they pertain to claims 1-5 and 19-20 are discussed in the 35 USC 103 rejection above.
Fuenzalida does not teach the mutation being a biallelic indel mutation.
Oji teaches wherein the mutation is a biallelic indel mutation (see pg. 1 “Here we show that large deletions with two sgRNAs as well as dsDNA-mediated point mutations are efficient in mouse embryonic stem cells (ESCs). The dsDNA-mediated gene knockins are also feasible in ESCs. Finally, we generated chimeric mice with biallelic mutant ESCs for a lethal gene, Dnajb13, and analyzed their phenotypes.”, see fig. 1) (instant claim 9).
It would have been obvious to one of ordinary skill in the art at the time the application
was filed to combine the Fuenzalida’s methods of selecting transformed cells with Pedersen’s teachings of using sgRNA for multiplex editing with Doench’s teachings of using Cas-9 and sgRNA’s from different constructs, and with Oji’s methods of biallelic genome editing. Oji teaches that most embryonic stem cell clones are biallelic mutant after sgRNA/CAS9-mediated genome editing, allowing for the ability to analyze the effects of genome editing by differentiating the mutant embryonic stem cells in vitro (see pg. 5). The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
3.Claim 7 is rejected under 35 U.S.C. 103 as being unpatentable over Fuenzalida,
Pedersen, and Doench as applied to claims 1-5 and 19-20 above, and in further view of Liu, Yongzhen et al. “CRISPR/Cas9-mediated p53 and Pten dual mutation accelerates hepatocarcinogenesis in adult hepatitis B virus transgenic mice.” Scientific reports vol. 7,1 2796. 5 Jun. 2017, doi:10.1038/s41598-017-03070-8.
The teachings of Fuenzalida, Pedersen, and Doench as they pertain to claims 1-5 and 19-20 are discussed in the 35 USC 103 rejection above.
Fuenzalida does not teach the target sequence being a nucleic acid sequence encoding PTEN, MYC, or ZMIZ1.
Liu teaches towards the target sequence being a nucleic acid sequence encoding PTEN (see pg. 2 “To induce p53 and Pten gene mutation simultaneously, we constructed a dual sgRNA cassette plasmid by inserting sgRNAs of p53 and Pten into the pSpCas(BB)-2A-GFP (PX458) vector. The sgRNAs specific for p53 or Pten were designed to target the first exon of mouse p53 or Pten gene, respectively.”) (instant claim 7).
It would have been obvious to one of ordinary skill in the art at the time the application
was filed to combine Fuenzalida’s methods of selecting transformed cells with Pedersen’s teachings of using sgRNA for multiplex editing with Doench’s teachings of using Cas-9 and sgRNA’s from different constructs, and with Liu’s teachings of using Pten as a target sequence. Liu teaches that p53 and altered Pten expression are the two most common genetic events in Hepatitis B virus infection related to hepatocellular carcinoma (HCC). Liu goes on to teach that in addition to p53 mutation, the inactivation of the phosphatase and tensin homolog (Pten) through genetic or post-translation modifications is found in about half of HCC patients (see pg. 1). Liu further teaches that liver-specific knockout of Pten in mice results in fatty liver disease and late-onset liver cancer (see pg.1). Lastly, Liu teaches that the loss of Pten function plays a pivotal role in promoting carcinogenesis of HCC (see pg. 1). The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
4.Claim 21 is rejected under 35 U.S.C. 103 as being unpatentable over Fuenzalida,
Pedersen, and Doench as applied to claims 1-5 and 19-20 above, and in further view of
Meissner, Torsten B et al. “Genome editing for human gene therapy.” Methods in enzymology
vol. 546 (2014): 273-95. doi:10.1016/B978-0-12-801185-0.00013-1 (List of references cited on 08/27/2024).
The teachings of Fuenzalida, Pedersen, and Doench as they pertain to claims 1-5 and 19-20 are discussed in the 35 USC 103 rejection above.
Fuenzalida does not teach the antibody being an anti-B2M antibody.
Meissner teaches wherein the antibody is an anti-B2M antibody (see pg. 276 “In the case of B2M, which is expressed at the cell surface, successful targeting can be confirmed by monitoring loss of B2M expression by means of a simple surface staining with an anti-B2M antibody, followed by fluorescence-activated cell sorting (FACS) analysis.”) (instant claim 21).
It would have been obvious to one of ordinary skill in the art at the time the application
was filed to combine Fuenzalida’s methods of selecting transformed cells with Pedersen’s teachings of using sgRNA for multiplex editing with Doench’s teachings of using Cas-9 and sgRNA’s from different constructs, with Meissner’s teachings of using an anti-B2M antibody for genome editing. Meissner teaches that the use of a surface antigen (B2M) allows the detection of successful targeting with a simple FACs staining using a fluorescently labeled antibody against the targeted protein (see pg. 282). The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
Response to Arguments
The arguments filed on 06/01/2026 have been considered by the examiner.
A new 35 USC 103 was necessitated by the amendments.
Applicants’ arguments were aimed at the previously now moot rejections. However, where relevant, applicant’s arguments are addressed in light of the new 103 rejections.
On p. 6 applicant argues that Fuenzalida and Pedersen do not teach the new limitations of “wherein the sgRNA against a marker gene targets an endogenous marker” and “wherein Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed from different constructs”.
However, Doench teaches the sgRNA against a marker gene targets an endogenous marker by teaching the sgRNA against a marker gene targeting CD13 and CD33, which are known endogenous markers in the art, and the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed on different constructs (see abstract, see page 1264, see figure 2). Fuenzalida teaches CRISPR consists of two components: a “guide” RNA (gRNA) and a non-specific CRISPR-associated endonuclease (Cas9) (see [0048]), applying CRISPR Cas-9 to a group (population) of cells and the population of cells consist of eukaryotic cells (see claims 1, 12, 19 and 22). Petersen teaches using sgRNA against a marker gene (see page 98 lines 1-5), and using FACs based negative selection of a population of cells (see claim 13).
On pp. 6-7 applicant argues that regardless if Oji teaches a biallelic mutations, Oji fails to teach the newly amended limitations of “wherein the sgRNA against a marker gene targets an endogenous marker” and “wherein Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed from different constructs”.
However, Doench teaches the sgRNA against a marker gene targets an endogenous marker by teaching the sgRNA against a marker gene targeting CD13 and CD33, which are known endogenous markers in the art. Doench further teaches the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed on different constructs, as discussed above.
Oji teaches biallelic indel mutations (see page 1 “Here we show that large deletions with two sgRNAs as well as dsDNA-mediated point mutations are efficient in mouse embryonic stem cells (ESCs). The dsDNA-mediated gene knockins are also feasible in ESCs. Finally, we generated chimeric mice with biallelic mutant ESCs for a lethal gene, Dnajb13, and analyzed their phenotypes.”, see fig. 1).
On pp. 7-8 applicant argues that Doench at best teaches using sgRNAs to target cell surface markers, which are the target sequence. Applicant argues that Doench does not teach sgRNA to target an endogenous marker and/or Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence being expressed from different constructs.
However, Doench explicitly teaches sgRNAs against six endogenous targets (see abstract “We created a pool of sgRNAs, tiling across all possible target sites of a panel of six endogenous mouse and three endogenous human genes and quantitatively assessed their ability to produce null alleles of their target gene by antibody staining and flow cytometry.”, see figure 1, see page 1268 under “FACs”).
Doench also teaches Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence being expressed from different constructs (see page 1268 “Cas9 activity assay. Cas9 expressing MOLM13, NB4 and TF1 cell lines were transduced with pXPR-011 (Addgene plasmid 59702) at an MOI ~1. Briefly, cells were infected in 24-well plate format, with each well containing 2 × 105 cells, 100 μl virus and 300 μl of media supplemented with 4 μg/ml polybrene. 48 h after infection, 2 μg/ml puromycin was added and cells were selected for 3 d. Parental lines transduced with pXPR-011 only were maintained in parallel with Cas9 and pXPR-011 expressing cell lines; samples from both were ana-lyzed on a BD-LSRFortessa X-20 ten d after infection. Active Cas9-expressing lines will result in a reduction in GFP when transfected with pXPR-011 as this vector delivers both GFP and a sgRNA targeting GFP. Because GFP is downstream of puromycin and after a 2A site, abrogation of GFP will have no impact on puromycin resistance (Supplementary Fig. 2)”, see pages 1263-1264 “…three human acute myeloid leukemia cell lines, MOLM13, NB4 and TF1, by transduction with a vector delivering Cas9 and conferring blasticidin resistance. We confirmed Cas9 activity in these polyclonal lines (Supplementary Fig. 2), then transduced each with the human sgRNA pool and collected marker-negative populations 8 d after transduction. CD15, CD13 and CD33 were evaluated in one, two and three of the cell lines, respectively. For all 12 sorted cell popula-tions, PCR of genomic DNA, followed by next-generation sequenc-ing, identified the sgRNAs that led to complete loss of the protein of interest (Supplementary Tables 2 and 3). We first examined the knockout specificity of the sgRNAs targeting each gene. In the mouse pool we observed 61–157 sgRNAs per gene enriched at least tenfold in each of the marker-negative cell popula-tions after normalizing the abundance of sgRNAs in each sorted pop-ulation to their starting abundance in the unsorted population. In the human pool, 116–256 sgRNAs per gene were at least twofold enriched after similar normalization; a lower threshold was used for this pool because each gene comprises a larger fraction of the overall pool…”).
On p. 9 applicant argues that regardless of whether Liu teaches PTEN, Liu fails to teach or suggest sgRNA to target an endogenous marker and/or Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence being expressed from different constructs.
However, Doench teaches the sgRNA against a marker gene targets an endogenous marker by teaching the sgRNA against a marker gene targeting CD13 and CD33, which are known endogenous markers in the art. Doench further teaches the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed on different constructs, as discussed above.
Liu teaches the target sequence being a nucleic acid sequence encoding PTEN (see page 2).
On pp. 9-10 applicant argues that Meissner fails to teach sgRNA to target an endogenous marker and/or Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence being expressed from different constructs.
However, Doench teaches the sgRNA against a marker gene targets an endogenous marker by teaching the sgRNA against a marker gene targeting CD13 and CD33, which are known endogenous markers in the art. Doench further teaches the Cas-9, sgRNA against a marker gene, and sgRNA against a target sequence are expressed on different constructs, as discussed above.
Meissner teaches wherein the antibody is an anti-B2M antibody (see page 276).
Allowable Subject Matter
Claims 6 and 8 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 prior art does not teach or suggest administering CAS-9, a sgRNA against a marker
gene, where the sgRNA against a marker gene comprises SEQ ID NO: 1, SEQ ID NO: 2, or
SEQ ID NO: 3 (instant claim 6), and an sgRNA against a target sequence to a population of
cells, where the sgRNA against a target sequence comprises SEQ ID NO: 7, SEQ ID NO: 8, or
SEQ ID NO: 9 (instant claim 8).
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). 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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/MCKENZIE A DUNN/Examiner, Art Unit 1678
/GREGORY S EMCH/Supervisory Patent Examiner, Art Unit 1678
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