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 .
Status of Claims
The response to the restriction requirement filed 02/20/2026 is acknowledged. Applicant elects Group I (claims 2, 53, 119, 133, 136, 137, 148, 151, 152, 163, 164, 167, 168, 172, 173, 185, 186, 620, 622, 624, 626, 628, 630, 632, 634, and 637; a composition and a cell) without traverse. The amendment filed 02/11/2026 is acknowledged. Claims 1 and 3-647 are cancelled. Claims 648-714 are new. Newly added claims 657-714 are distinct from the elected group and are therefore withdrawn and not under examination. Claims 2 and 648-656 are pending and examined.
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 652-655 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. The phrases “in combination with” and “associated with” in claims 652 and 655 render the claims indefinite because neither the claims nor the specification defines what it means to be “in combination” or “associated.” It is unclear whether the components are required to be physically associated or combined, co-formulated, co-administered, or together in a different sense. Accordingly, a person having ordinary skill in the art would not be able to determine the metes and bounds of the claim with reasonable certainty. Claims 653 and 654 are also rejected for depending upon an indefinite claim.
Therefore, claims 652-655 are rejected under 35 U.S.C. 112(b) for being indefinite.
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.
Claims 2, 649-651, and 656 are rejected under 35 U.S.C. 103 as being unpatentable over Baik et al. (US Patent 11208458), in view of Anguela et al. (WO2019222411A1).
Claim 2 is drawn to a nucleic acid encoding a multidomain therapeutic protein, claim 649 is drawn to specific structural features, claims 650 and 651 are drawn to a viral vector comprising the nucleic acid, claim 656 is drawn to a cell comprising the nucleic acid.
Baik et al. discloses a polynucleotide encoding a multidomain therapeutic protein comprising a delivery domain and an enzyme domain, wherein the delivery domain binds to CD63 [see claim 1] (instant claim 2) and the enzyme domain comprises an alpha-glucosidase [see claim 5] (instant claim 2). Baik et al. utilizes the terms alpha glucosidase and lysosomal alpha glucosidase synonymously [see specification, col. 3, par. 1, lines 3-4 and col. 5, par. 5, lines 33-35], and as such, one would reasonably believe that claims recite lysosomal alpha-glucosidase (instant claim 2). Additionally, the specification notes that the term “polynucleotide” encompasses DNA and RNA [see specification, col. 32, par. 6, lines 52-59] (instant claim 650). This polynucleotide lacks a homology arm, as evidenced by dependent claim 7 disclosing an embodiment in which the polynucleotide further comprises a locus-targeting sequence [see claim 7], which the specification defines to include flanking homology arms that enable homologous recombination [see specification, col. 34, par. 1, lines 5-7] (instant claim 649). Further still, it would have been obvious to a person having ordinary skill in the art that not all embodiments require flanking homology arms as homologous recombination is not necessary, for example, when only transient expression is required. Baik et al. discloses that the locus targeting sequence may also comprise a guide RNA sequence, which inherently comprises a DNA targeting segment (instant claim 653) and a type II Cas enzyme (which comprises Cas9 endonucleases), describing this as “the CRISPR-Cas9 method” [see specification, col. 34, lines 7-10] (instant claims 652 and 653). The co-expression of these two compositions allows the guide RNA to facilitate Cas9 targeting of the nuclease site (instant claim 652 and 653). The locus targeting sequence may target a human albumin locus [see claim 8] (claim 652). Baik et al. discloses the composition in an adeno-associated virus (AAV) [see claim 8] (instant claim 650), which includes AAV8 [see specification, col. 35, par. 3, lines 31-34], and because it was known in the art that AAV8 is naturally single stranded, a person having ordinary skill in the art would reasonably believe that the disclosed AAV8 would be single stranded unless otherwise stated (instant claim 651). Baik et al. discloses that the polynucleotide sequence encoding the multidomain therapeutic protein of claim 1 encodes the amino acid sequence of SEQ ID NO: 79 [see claim 9], which corresponds to instant SEQ ID NO: 193 [see alignment below] (instant claim 648).
SEQ ID NO: 79 aligned to instant SEQ ID NO: 193
US-16-001-717-79
Sequence 79, US/16001717
Publication No. US20180355017A1
Query Match 100.0%; Score 6042; Length 1128;
Best Local Similarity 100.0%;
Matches 1128; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLQESGPKVVKPSETLSLTCTVSGGSISSYYWNWIRQSPGKGLEWIGYTKRGYTDYNP 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLQESGPKVVKPSETLSLTCTVSGGSISSYYWNWIRQSPGKGLEWIGYTKRGYTDYNP 60
Qy 61 SLRSRVTISEDTSKNQFSLRISSVTAADTAVYYCAQMGWGSHAFDMWGQGTMVAVSSGGS 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 SLRSRVTISEDTSKNQFSLRISSVTAADTAVYYCAQMGWGSHAFDMWGQGTMVAVSSGGS 120
Qy 121 GGGGSGGGGSGGEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLL 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GGGGSGGGGSGGEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLL 180
Qy 181 IYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 IYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK 240
Qy 241 GGGGSAHPGRPRAVPTQCDVPPNSRFDCAPDKAITQEQCEARGCCYIPAKQGLQGAQMGQ 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GGGGSAHPGRPRAVPTQCDVPPNSRFDCAPDKAITQEQCEARGCCYIPAKQGLQGAQMGQ 300
Qy 301 PWCFFPPSYPSYKLENLSSSEMGYTATLTRTTPTFFPKDILTLRLDVMMETENRLHFTIK 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 PWCFFPPSYPSYKLENLSSSEMGYTATLTRTTPTFFPKDILTLRLDVMMETENRLHFTIK 360
Qy 361 DPANRRYEVPLETPHVHSRAPSPLYSVEFSEEPFGVIVRRQLDGRVLLNTTVAPLFFADQ 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 DPANRRYEVPLETPHVHSRAPSPLYSVEFSEEPFGVIVRRQLDGRVLLNTTVAPLFFADQ 420
Qy 421 FLQLSTSLPSQYITGLAEHLSPLMLSTSWTRITLWNRDLAPTPGANLYGSHPFYLALEDG 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 FLQLSTSLPSQYITGLAEHLSPLMLSTSWTRITLWNRDLAPTPGANLYGSHPFYLALEDG 480
Qy 481 GSAHGVFLLNSNAMDVVLQPSPALSWRSTGGILDVYIFLGPEPKSVVQQYLDVVGYPFMP 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GSAHGVFLLNSNAMDVVLQPSPALSWRSTGGILDVYIFLGPEPKSVVQQYLDVVGYPFMP 540
Qy 541 PYWGLGFHLCRWGYSSTAITRQVVENMTRAHFPLDVQWNDLDYMDSRRDFTFNKDGFRDF 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 PYWGLGFHLCRWGYSSTAITRQVVENMTRAHFPLDVQWNDLDYMDSRRDFTFNKDGFRDF 600
Qy 601 PAMVQELHQGGRRYMMIVDPAISSSGPAGSYRPYDEGLRRGVFITNETGQPLIGKVWPGS 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 PAMVQELHQGGRRYMMIVDPAISSSGPAGSYRPYDEGLRRGVFITNETGQPLIGKVWPGS 660
Qy 661 TAFPDFTNPTALAWWEDMVAEFHDQVPFDGMWIDMNEPSNFIRGSEDGCPNNELENPPYV 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 TAFPDFTNPTALAWWEDMVAEFHDQVPFDGMWIDMNEPSNFIRGSEDGCPNNELENPPYV 720
Qy 721 PGVVGGTLQAATICASSHQFLSTHYNLHNLYGLTEAIA SHRALVKARGTRPFVISRSTFA 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 PGVVGGTLQAATICASSHQFLSTHYNLHNLYGLTEAIA SHRALVKARGTRPFVISRSTFA 780
Qy 781 GHGRYAGHWTGDVWSSWEQLASSVPEILQFNLLGVPLVGADVCGFLGNTSEELCVRWTQL 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GHGRYAGHWTGDVWSSWEQLASSVPEILQFNLLGVPLVGADVCGFLGNTSEELCVRWTQL 840
Qy 841 GAFYPFMRNHNSLLSLPQEPYSFSEPAQQAMRKALTLRYALLPHLYTLFHQAHVAGETVA 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 GAFYPFMRNHNSLLSLPQEPYSFSEPAQQAMRKALTLRYALLPHLYTLFHQAHVAGETVA 900
Qy 901 RPLFLEFPKDSSTWTVDHQLLWGEALLITPVLQAGKAEVTGYFPLGTWYDLQTVPVEALG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 RPLFLEFPKDSSTWTVDHQLLWGEALLITPVLQAGKAEVTGYFPLGTWYDLQTVPVEALG 960
Qy 961 SLPPPPAAPREPAIHSEGQWVTLPAPLDTINVHLRAGYIIPLQGPGLTTTESRQQPMALA 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 SLPPPPAAPREPAIHSEGQWVTLPAPLDTINVHLRAGYIIPLQGPGLTTTESRQQPMALA 1020
Qy 1021 VALTKGGEARGELFWDDGESLEVLERGAYTQVIFLARNNTIVNELVRVTSEGAGLQLQKV 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 VALTKGGEARGELFWDDGESLEVLERGAYTQVIFLARNNTIVNELVRVTSEGAGLQLQKV 1080
Qy 1081 TVLGVATAPQQVLSNGVPVSNFTYSPDTKVLDICVSLLMGEQFLVSWC 1128
||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 TVLGVATAPQQVLSNGVPVSNFTYSPDTKVLDICVSLLMGEQFLVSWC 1128
Baik et al. discloses a method of expressing a multidomain therapeutic protein in a cell, comprising contacting a cell with a vector comprising the polynucleotide discussed above and allowing the polynucleotide to integrate into a genomic locus [see claims 1, 10, and 16]. For the polynucleotide to integrate it must necessarily be inside of the cell (instant claim 656).
Baik et al. does not disclose that the polynucleotide is CpG depleted. Of note, the instant application defines CpG depletion as having one or more CpG dinucleotides removed [see instant specification, par. 265, lines 5-6].
Anguela et al. discloses a nucleic acid encoding an acid a-glucosidase wherein the nucleic acid has no CpG dinucleotides [see claims 1 and 32] compared to a reference nucleic acid [see par. 127]. Acid a-glucosidase is known in the art as an alternative name of lysosomal alpha-glucosidase. Anguela et al. discloses that removal of CpG, or in other words CpG depletion, is a method of optimizing codons to improve protein expression [see p. 1, par. 4, lines 1-3 and p. 79, par. 416, lines 4-8].
It would have been obvious to combine these teachings and remove the CpG dinucleotides from the composition of Baik et al. as the technique was a known method of increasing protein expression, and both are directed to alpha-glucosidase (instant claim 2).
Therefore, claims 2, 649-651, and 656 are rejected under 35 U.S.C. 103.
Claims 652 and 653 are rejected under 35 U.S.C. 103 as being unpatentable over Baik et al. (US Patent 11208458) in view of Anguela et al. (WO2019222411A1) as applied to claims 2, 649-651, and 656 above, and further in view of Holmes et al. (US Patent 10639383).
Claims 652 and 653 are drawn to specific features of a nuclease agent.
The disclosure of Baik et al. and Anguela et al. are discussed above.
The combination of these disclosures does not teach or suggest a specific DNA targeted segment.
Holmes et al. discloses a method of expressing an antibody in a liver cell, comprising administering an AAV comprising a transgene and an additional AAV comprising a nuclease that cleaves an endogenous albumin gene in a liver cell and allows integration of the transgene [see claim 1]. To this end, Holmes et al. discloses CRISPR/Cas9 systems targeting the human albumin locus for therapeutic genomic integration in liver cells, and expressly discloses intron 1 [see col. 38, lines 46-49] (instant claim 652) albumin-targeted guide RNAs (labeled as gRNA) [see col. 39, table 3]. Table 3 discloses guide RNA Alb-r1469, which consists of the guide RNA’s target sequence and corresponding guide RNA sequence; the guide RNA sequence is enumerated in SEQ ID NO: 22 corresponds to instant SEQ ID NO: 36 [see alignment below] (instant claim 654).
SEQ ID NO: 22 aligned to instant SEQ ID NO: 36
US-15-357-772-22
Sequence 22, US/15357772
Publication No. US20170196992A1
Query Match 100.0%; Score 20; Length 21;
Best Local Similarity 100.0%;
Matches 20; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 TAAAGCATAGTGCAATGGAT 20
||||||||||||||||||||
Db 2 TAAAGCATAGTGCAATGGAT 21
Holmes et al. further demonstrates successful cleavage activity of the Alb-r1469 system in human cells [see col. 40, table 4].
It would have been obvious to a person having ordinary skill in the art to combine these teachings and utilize the albumin-targeting guide RNA sequence taught by Holmes et al. because Holmes et al. discloses that the recited guide RNA effectively targets and cleaves the human albumin locus. Furthermore, one would have reasonably expected success in combining these teachings because both references are directed to liver-targeted therapeutic genomic integration at the albumin safe harbor locus using CRISPR/Cas9.
Therefore, claims 652 and 653 are rejected under 35 U.S.C. 103.
Claims 654 and 655 are rejected under 35 U.S.C. 103 as being unpatentable over Baik et al. (US Patent 11208458) in view of Anguela et al. (WO2019222411A1) and Holmes et al. (US Patent 10639383) as applied to claims 2, 649-653, and 656 above, and further in view of Dombrowski et al. (US Patent 11697806).
Claim 654 is drawn to specific features of the guide RNA and Cas9; claim 655 is drawn to specific features of a lipid nanoparticle comprising the Cas9 and RNA.
The disclosure of Baik et al., Anguela et al., and Holmes et al. are discussed above.
The combination of these disclosures does not teach or suggest a specific sequence for the Cas9 protein or lipid nanoparticles.
Dombrowski et al. discloses an mRNA comprising a Cas9 open reading frame whose sequence is enumerated in SEQ ID NO: 111. This sequence corresponds to instant SEQ ID NO: 2 [see alignment below] (instant claim 654).
SEQ ID NO: 111 aligned to instant SEQ ID NO: 2
US-16-828-615A-111
Sequence 111, US/16828615A
Publication No. US20200354702A1
Query Match 100.0%; Score 4140; Length 4140;
Best Local Similarity 100.0%;
Matches 4140; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATGGACAAGAAGTACTCCATCGGCCTGGACATCGGCACCAACTCCGTGGGCTGGGCCGTG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ATGGACAAGAAGTACTCCATCGGCCTGGACATCGGCACCAACTCCGTGGGCTGGGCCGTG 60
Qy 61 ATCACCGACGAGTACAAGGTGCCCTCCAAGAAGTTCAAGGTGCTGGGCAACACCGACCGG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ATCACCGACGAGTACAAGGTGCCCTCCAAGAAGTTCAAGGTGCTGGGCAACACCGACCGG 120
Qy 121 CACTCCATCAAGAAGAACCTGATCGGCGCCCTGCTGTTCGACTCCGGCGAGACCGCCGAG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 CACTCCATCAAGAAGAACCTGATCGGCGCCCTGCTGTTCGACTCCGGCGAGACCGCCGAG 180
Qy 181 GCCACCCGGCTGAAGCGGACCGCCCGGCGGCGGTACACCCGGCGGAAGAACCGGATCTGC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GCCACCCGGCTGAAGCGGACCGCCCGGCGGCGGTACACCCGGCGGAAGAACCGGATCTGC 240
Qy 241 TACCTGCAGGAGATCTTCTCCAACGAGATGGCCAAGGTGGACGACTCCTTCTTCCACCGG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TACCTGCAGGAGATCTTCTCCAACGAGATGGCCAAGGTGGACGACTCCTTCTTCCACCGG 300
Qy 301 CTGGAGGAGTCCTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGGCACCCCATCTTCGGC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 CTGGAGGAGTCCTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGGCACCCCATCTTCGGC 360
Qy 361 AACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGCGGAAG 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 AACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGCGGAAG 420
Qy 421 AAGCTGGTGGACTCCACCGACAAGGCCGACCTGCGGCTGATCTACCTGGCCCTGGCCCAC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 AAGCTGGTGGACTCCACCGACAAGGCCGACCTGCGGCTGATCTACCTGGCCCTGGCCCAC 480
Qy 481 ATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACTCCGAC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACTCCGAC 540
Qy 541 GTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAGAACCCC 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 GTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAGAACCCC 600
Qy 601 ATCAACGCCTCCGGCGTGGACGCCAAGGCCATCCTGTCCGCCCGGCTGTCCAAGTCCCGG 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ATCAACGCCTCCGGCGTGGACGCCAAGGCCATCCTGTCCGCCCGGCTGTCCAAGTCCCGG 660
Qy 661 CGGCTGGAGAACCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAACGGCCTGTTCGGCAAC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 CGGCTGGAGAACCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAACGGCCTGTTCGGCAAC 720
Qy 721 CTGATCGCCCTGTCCCTGGGCCTGACCCCCAACTTCAAGTCCAACTTCGACCTGGCCGAG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 CTGATCGCCCTGTCCCTGGGCCTGACCCCCAACTTCAAGTCCAACTTCGACCTGGCCGAG 780
Qy 781 GACGCCAAGCTGCAGCTGTCCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCC 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GACGCCAAGCTGCAGCTGTCCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCC 840
Qy 841 CAGATCGGCGACCAGTACGCCGACCTGTTCCTGGCCGCCAAGAACCTGTCCGACGCCATC 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 CAGATCGGCGACCAGTACGCCGACCTGTTCCTGGCCGCCAAGAACCTGTCCGACGCCATC 900
Qy 901 CTGCTGTCCGACATCCTGCGGGTGAACACCGAGATCACCAAGGCCCCCCTGTCCGCCTCC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 CTGCTGTCCGACATCCTGCGGGTGAACACCGAGATCACCAAGGCCCCCCTGTCCGCCTCC 960
Qy 961 ATGATCAAGCGGTACGACGAGCACCACCAGGACCTGACCCTGCTGAAGGCCCTGGTGCGG 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 ATGATCAAGCGGTACGACGAGCACCACCAGGACCTGACCCTGCTGAAGGCCCTGGTGCGG 1020
Qy 1021 CAGCAGCTGCCCGAGAAGTACAAGGAGATCTTCTTCGACCAGTCCAAGAACGGCTACGCC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 CAGCAGCTGCCCGAGAAGTACAAGGAGATCTTCTTCGACCAGTCCAAGAACGGCTACGCC 1080
Qy 1081 GGCTACATCGACGGCGGCGCCTCCCAGGAGGAGTTCTACAAGTTCATCAAGCCCATCCTG 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 GGCTACATCGACGGCGGCGCCTCCCAGGAGGAGTTCTACAAGTTCATCAAGCCCATCCTG 1140
Qy 1141 GAGAAGATGGACGGCACCGAGGAGCTGCTGGTGAAGCTGAACCGGGAGGACCTGCTGCGG 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 GAGAAGATGGACGGCACCGAGGAGCTGCTGGTGAAGCTGAACCGGGAGGACCTGCTGCGG 1200
Qy 1201 AAGCAGCGGACCTTCGACAACGGCTCCATCCCCCACCAGATCCACCTGGGCGAGCTGCAC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 AAGCAGCGGACCTTCGACAACGGCTCCATCCCCCACCAGATCCACCTGGGCGAGCTGCAC 1260
Qy 1261 GCCATCCTGCGGCGGCAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGGGAGAAGATC 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 GCCATCCTGCGGCGGCAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGGGAGAAGATC 1320
Qy 1321 GAGAAGATCCTGACCTTCCGGATCCCCTACTACGTGGGCCCCCTGGCCCGGGGCAACTCC 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 GAGAAGATCCTGACCTTCCGGATCCCCTACTACGTGGGCCCCCTGGCCCGGGGCAACTCC 1380
Qy 1381 CGGTTCGCCTGGATGACCCGGAAGTCCGAGGAGACCATCACCCCCTGGAACTTCGAGGAG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1381 CGGTTCGCCTGGATGACCCGGAAGTCCGAGGAGACCATCACCCCCTGGAACTTCGAGGAG 1440
Qy 1441 GTGGTGGACAAGGGCGCCTCCGCCCAGTCCTTCATCGAGCGGATGACCAACTTCGACAAG 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1441 GTGGTGGACAAGGGCGCCTCCGCCCAGTCCTTCATCGAGCGGATGACCAACTTCGACAAG 1500
Qy 1501 AACCTGCCCAACGAGAAGGTGCTGCCCAAGCACTCCCTGCTGTACGAGTACTTCACCGTG 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 AACCTGCCCAACGAGAAGGTGCTGCCCAAGCACTCCCTGCTGTACGAGTACTTCACCGTG 1560
Qy 1561 TACAACGAGCTGACCAAGGTGAAGTACGTGACCGAGGGCATGCGGAAGCCCGCCTTCCTG 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1561 TACAACGAGCTGACCAAGGTGAAGTACGTGACCGAGGGCATGCGGAAGCCCGCCTTCCTG 1620
Qy 1621 TCCGGCGAGCAGAAGAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAGGTGACC 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1621 TCCGGCGAGCAGAAGAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAGGTGACC 1680
Qy 1681 GTGAAGCAGCTGAAGGAGGACTACTTCAAGAAGATCGAGTGCTTCGACTCCGTGGAGATC 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1681 GTGAAGCAGCTGAAGGAGGACTACTTCAAGAAGATCGAGTGCTTCGACTCCGTGGAGATC 1740
Qy 1741 TCCGGCGTGGAGGACCGGTTCAACGCCTCCCTGGGCACCTACCACGACCTGCTGAAGATC 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1741 TCCGGCGTGGAGGACCGGTTCAACGCCTCCCTGGGCACCTACCACGACCTGCTGAAGATC 1800
Qy 1801 ATCAAGGACAAGGACTTCCTGGACAACGAGGAGAACGAGGACATCCTGGAGGACATCGTG 1860
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1801 ATCAAGGACAAGGACTTCCTGGACAACGAGGAGAACGAGGACATCCTGGAGGACATCGTG 1860
Qy 1861 CTGACCCTGACCCTGTTCGAGGACCGGGAGATGATCGAGGAGCGGCTGAAGACCTACGCC 1920
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1861 CTGACCCTGACCCTGTTCGAGGACCGGGAGATGATCGAGGAGCGGCTGAAGACCTACGCC 1920
Qy 1921 CACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGGCGGCGGTACACCGGCTGGGGC 1980
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1921 CACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGGCGGCGGTACACCGGCTGGGGC 1980
Qy 1981 CGGCTGTCCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACCATCCTG 2040
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1981 CGGCTGTCCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACCATCCTG 2040
Qy 2041 GACTTCCTGAAGTCCGACGGCTTCGCCAACCGGAACTTCATGCAGCTGATCCACGACGAC 2100
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2041 GACTTCCTGAAGTCCGACGGCTTCGCCAACCGGAACTTCATGCAGCTGATCCACGACGAC 2100
Qy 2101 TCCCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTGTCCGGCCAGGGCGACTCCCTG 2160
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2101 TCCCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTGTCCGGCCAGGGCGACTCCCTG 2160
Qy 2161 CACGAGCACATCGCCAACCTGGCCGGCTCCCCCGCCATCAAGAAGGGCATCCTGCAGACC 2220
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2161 CACGAGCACATCGCCAACCTGGCCGGCTCCCCCGCCATCAAGAAGGGCATCCTGCAGACC 2220
Qy 2221 GTGAAGGTGGTGGACGAGCTGGTGAAGGTGATGGGCCGGCACAAGCCCGAGAACATCGTG 2280
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2221 GTGAAGGTGGTGGACGAGCTGGTGAAGGTGATGGGCCGGCACAAGCCCGAGAACATCGTG 2280
Qy 2281 ATCGAGATGGCCCGGGAGAACCAGACCACCCAGAAGGGCCAGAAGAACTCCCGGGAGCGG 2340
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2281 ATCGAGATGGCCCGGGAGAACCAGACCACCCAGAAGGGCCAGAAGAACTCCCGGGAGCGG 2340
Qy 2341 ATGAAGCGGATCGAGGAGGGCATCAAGGAGCTGGGCTCCCAGATCCTGAAGGAGCACCCC 2400
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2341 ATGAAGCGGATCGAGGAGGGCATCAAGGAGCTGGGCTCCCAGATCCTGAAGGAGCACCCC 2400
Qy 2401 GTGGAGAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAACGGCCGG 2460
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2401 GTGGAGAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAACGGCCGG 2460
Qy 2461 GACATGTACGTGGACCAGGAGCTGGACATCAACCGGCTGTCCGACTACGACGTGGACCAC 2520
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2461 GACATGTACGTGGACCAGGAGCTGGACATCAACCGGCTGTCCGACTACGACGTGGACCAC 2520
Qy 2521 ATCGTGCCCCAGTCCTTCCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCCGGTCC 2580
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2521 ATCGTGCCCCAGTCCTTCCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCCGGTCC 2580
Qy 2581 GACAAGAACCGGGGCAAGTCCGACAACGTGCCCTCCGAGGAGGTGGTGAAGAAGATGAAG 2640
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2581 GACAAGAACCGGGGCAAGTCCGACAACGTGCCCTCCGAGGAGGTGGTGAAGAAGATGAAG 2640
Qy 2641 AACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATCACCCAGCGGAAGTTCGACAACCTG 2700
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2641 AACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATCACCCAGCGGAAGTTCGACAACCTG 2700
Qy 2701 ACCAAGGCCGAGCGGGGCGGCCTGTCCGAGCTGGACAAGGCCGGCTTCATCAAGCGGCAG 2760
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2701 ACCAAGGCCGAGCGGGGCGGCCTGTCCGAGCTGGACAAGGCCGGCTTCATCAAGCGGCAG 2760
Qy 2761 CTGGTGGAGACCCGGCAGATCACCAAGCACGTGGCCCAGATCCTGGACTCCCGGATGAAC 2820
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2761 CTGGTGGAGACCCGGCAGATCACCAAGCACGTGGCCCAGATCCTGGACTCCCGGATGAAC 2820
Qy 2821 ACCAAGTACGACGAGAACGACAAGCTGATCCGGGAGGTGAAGGTGATCACCCTGAAGTCC 2880
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2821 ACCAAGTACGACGAGAACGACAAGCTGATCCGGGAGGTGAAGGTGATCACCCTGAAGTCC 2880
Qy 2881 AAGCTGGTGTCCGACTTCCGGAAGGACTTCCAGTTCTACAAGGTGCGGGAGATCAACAAC 2940
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2881 AAGCTGGTGTCCGACTTCCGGAAGGACTTCCAGTTCTACAAGGTGCGGGAGATCAACAAC 2940
Qy 2941 TACCACCACGCCCACGACGCCTACCTGAACGCCGTGGTGGGCACCGCCCTGATCAAGAAG 3000
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2941 TACCACCACGCCCACGACGCCTACCTGAACGCCGTGGTGGGCACCGCCCTGATCAAGAAG 3000
Qy 3001 TACCCCAAGCTGGAGTCCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 3060
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3001 TACCCCAAGCTGGAGTCCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 3060
Qy 3061 ATGATCGCCAAGTCCGAGCAGGAGATCGGCAAGGCCACCGCCAAGTACTTCTTCTACTCC 3120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3061 ATGATCGCCAAGTCCGAGCAGGAGATCGGCAAGGCCACCGCCAAGTACTTCTTCTACTCC 3120
Qy 3121 AACATCATGAACTTCTTCAAGACCGAGATCACCCTGGCCAACGGCGAGATCCGGAAGCGG 3180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3121 AACATCATGAACTTCTTCAAGACCGAGATCACCCTGGCCAACGGCGAGATCCGGAAGCGG 3180
Qy 3181 CCCCTGATCGAGACCAACGGCGAGACCGGCGAGATCGTGTGGGACAAGGGCCGGGACTTC 3240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3181 CCCCTGATCGAGACCAACGGCGAGACCGGCGAGATCGTGTGGGACAAGGGCCGGGACTTC 3240
Qy 3241 GCCACCGTGCGGAAGGTGCTGTCCATGCCCCAGGTGAACATCGTGAAGAAGACCGAGGTG 3300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3241 GCCACCGTGCGGAAGGTGCTGTCCATGCCCCAGGTGAACATCGTGAAGAAGACCGAGGTG 3300
Qy 3301 CAGACCGGCGGCTTCTCCAAGGAGTCCATCCTGCCCAAGCGGAACTCCGACAAGCTGATC 3360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3301 CAGACCGGCGGCTTCTCCAAGGAGTCCATCCTGCCCAAGCGGAACTCCGACAAGCTGATC 3360
Qy 3361 GCCCGGAAGAAGGACTGGGACCCCAAGAAGTACGGCGGCTTCGACTCCCCCACCGTGGCC 3420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3361 GCCCGGAAGAAGGACTGGGACCCCAAGAAGTACGGCGGCTTCGACTCCCCCACCGTGGCC 3420
Qy 3421 TACTCCGTGCTGGTGGTGGCCAAGGTGGAGAAGGGCAAGTCCAAGAAGCTGAAGTCCGTG 3480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3421 TACTCCGTGCTGGTGGTGGCCAAGGTGGAGAAGGGCAAGTCCAAGAAGCTGAAGTCCGTG 3480
Qy 3481 AAGGAGCTGCTGGGCATCACCATCATGGAGCGGTCCTCCTTCGAGAAGAACCCCATCGAC 3540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3481 AAGGAGCTGCTGGGCATCACCATCATGGAGCGGTCCTCCTTCGAGAAGAACCCCATCGAC 3540
Qy 3541 TTCCTGGAGGCCAAGGGCTACAAGGAGGTGAAGAAGGACCTGATCATCAAGCTGCCCAAG 3600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3541 TTCCTGGAGGCCAAGGGCTACAAGGAGGTGAAGAAGGACCTGATCATCAAGCTGCCCAAG 3600
Qy 3601 TACTCCCTGTTCGAGCTGGAGAACGGCCGGAAGCGGATGCTGGCCTCCGCCGGCGAGCTG 3660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3601 TACTCCCTGTTCGAGCTGGAGAACGGCCGGAAGCGGATGCTGGCCTCCGCCGGCGAGCTG 3660
Qy 3661 CAGAAGGGCAACGAGCTGGCCCTGCCCTCCAAGTACGTGAACTTCCTGTACCTGGCCTCC 3720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3661 CAGAAGGGCAACGAGCTGGCCCTGCCCTCCAAGTACGTGAACTTCCTGTACCTGGCCTCC 3720
Qy 3721 CACTACGAGAAGCTGAAGGGCTCCCCCGAGGACAACGAGCAGAAGCAGCTGTTCGTGGAG 3780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3721 CACTACGAGAAGCTGAAGGGCTCCCCCGAGGACAACGAGCAGAAGCAGCTGTTCGTGGAG 3780
Qy 3781 CAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCTCCGAGTTCTCCAAGCGGGTG 3840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3781 CAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCTCCGAGTTCTCCAAGCGGGTG 3840
Qy 3841 ATCCTGGCCGACGCCAACCTGGACAAGGTGCTGTCCGCCTACAACAAGCACCGGGACAAG 3900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3841 ATCCTGGCCGACGCCAACCTGGACAAGGTGCTGTCCGCCTACAACAAGCACCGGGACAAG 3900
Qy 3901 CCCATCCGGGAGCAGGCCGAGAACATCATCCACCTGTTCACCCTGACCAACCTGGGCGCC 3960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3901 CCCATCCGGGAGCAGGCCGAGAACATCATCCACCTGTTCACCCTGACCAACCTGGGCGCC 3960
Qy 3961 CCCGCCGCCTTCAAGTACTTCGACACCACCATCGACCGGAAGCGGTACACCTCCACCAAG 4020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3961 CCCGCCGCCTTCAAGTACTTCGACACCACCATCGACCGGAAGCGGTACACCTCCACCAAG 4020
Qy 4021 GAGGTGCTGGACGCCACCCTGATCCACCAGTCCATCACCGGCCTGTACGAGACCCGGATC 4080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4021 GAGGTGCTGGACGCCACCCTGATCCACCAGTCCATCACCGGCCTGTACGAGACCCGGATC 4080
Qy 4081 GACCTGTCCCAGCTGGGCGGCGACGGCGGCGGCTCCCCCAAGAAGAAGCGGAAGGTGTGA 4140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4081 GACCTGTCCCAGCTGGGCGGCGACGGCGGCGGCTCCCCCAAGAAGAAGCGGAAGGTGTGA 4140
Dombrowski et al. discloses an embodiment in which the mRNA is inside of a lipid nanoparticle and comprises a 5’ cap, an n1-methyl-pseudouridine at all positions [see specification, col. 3-5, embodiments 1, 24, 64, 67, 70, 72 and 88] (instant claim 654 and 655). Dombrowski et al. discloses that substitution of N1- methyl-pseudouridine triggers less cytokine induction than unmodified Cas9 [see col. 76, par. 6, lines 35-40]. The Cas9 mRNA may comprise a polyA tail [see col. 94, par. 1, lines 11-14], which the instant specification uses interchangeably with the terms “polyadenylation sequence” and “polyadenylation tail sequence” [see instant specification par. 247] (instant claim 654). A suitable neutral lipid includes distearoylphosphatidylcholine (DSPC) [see col. 55, par. 5., lines 55-62] (instant claim 655).
It would have been obvious to combine these disclosures and utilize the guide RNA disclosed by Holmes et al. with the Cas9 disclosed by Dombrowski et al. as the art teaches that, in gene editing, unwanted elevations in cytokines may occur [see specification, col. 1, par 4, lines 26-29] and that this modified Cas9 produces less cytokine response. One would have reasonably expected that the modified Cas9 protein would function with the disclosed guide RNA of Holmes as both are directed to liver-targeted therapeutic genomic integration at the albumin locus using CRISPR/Cas9 and were demonstrated to work in similar systems.
Therefore, claims 654 and 655 are rejected under 35 U.S.C. 103.
Allowable Subject Matter
Claim 648 is 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: Although nucleic acid constructs encoding multidomain therapeutic proteins that comprise CD63 and lysosomal alpha glucosidase are known in the art, the prior art fails to teach or suggest a nucleic acid construct as set forth in SEQ ID NO: 196.
The closest prior art is Baik et al. (US Patent 11208458), which teaches a nucleic acid construct encoding a multidomain therapeutic protein comprising the amino acid sequence of instant SEQ ID NO: 193. However, it has been determined that the prior art does not teach the claimed nucleic acid construct with CpG depletion as enumerated in the distinct nucleic acid sequence of SEQ ID NO 196. The claimed nucleic acid construct is therefore free of the art.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Tirone D Johnson whose telephone number is (571)272-1256. The examiner can normally be reached M-F, 9-5 ET.
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/TIRONE D. JOHNSON/ Examiner, Art Unit 1675
/JEFFREY STUCKER/ Supervisory Patent Examiner, Art Unit 1675
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