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 .
Detailed Action
This action is in response to the papers filed March 30, 2026.
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 03/30/2026 has been entered.
Claim Amendments
Applicant’s amendment to the claims filed 03/30/2026 is acknowledged.
Claims 2, 12-13, 21-26 have been cancelled.
Claims 1 and 14 are amended.
Claims 1, 3-11, 14-20, 27-28 are pending.
Claims 4-5, 7-11, 16-20 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species/invention.
Claims 1, 3, 6, 14-15, 27-28 are under examination.
Election/Restrictions
The following is a summary of the restriction/election requirements in the application. See Requirement for Restriction/Election mailed 10/13/2023.
In the reply filed 12/12/2023, applicant elected without traverse:
the species of introducing foreign DNA into a targeted genome with homologous recombination of one of a 5'-end or a 3'-end of the foreign DNA and non-homologous recombination of the other end when double-strand breaks of a targeted genomic DNA occur, as the means of introducing foreign DNA into a genome; and
the species of a T cell, as the cell type that is genetically modified.
Accordingly, claims 4-5, 7-11, 16-20 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 12/12/2023.
Priority
The instant application 17/042,819 was filed on 09/28/2020. This application is a national stage of international application PCT/JP2019/013602 filed 03/28/2019, claiming priority based on Japanese patent application JP2018-066174 filed 03/29/2018.
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55. While a certified copy of the foreign patent application is provided with the instant application, a certified English translation of said foreign patent application has not been provided.
Withdrawal of Prior Rejections/Objections
Rejections and/or objections not reiterated from the previous Office action mailed 12/01/2025 are hereby withdrawn. The following rejections and/or objections are either newly applied or are reiterated and are the only rejections and/or objections presently applied to the instant application. Applicant’s remarks filed 03/30/2026 have been carefully considered but are moot in view of the new grounds of rejection set forth in this action.
Claim Interpretation
Page 21, lines 2-3, of the specification defines the claim term “non-homologous recombination” as meaning “non-homologous end joining.”
Allowable Subject Matter
Allowable subject matter has been communicated in the interview summary dated 04/21/2026.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 6, 14-15 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Lu et al. (2018) “Single-homology-arm linear DNA recombination by the nonhomologous end joining pathway as a novel and simple gene inactivation method: a proof-of-concept study in Dietzia sp. strain DQ12-45-1b” Applied and Environmental Microbiology, 84(19), e00795-18, 16 pages.
This rejection is newly applied, necessitated by amendment.
Claims 1 and 14 are directed to methods comprising a step (i.e., manipulative action) of introducing foreign DNA comprising a 5’-end and a 3’-end into a targeted genome of a cell, wherein the foreign DNA does not have a homology arm at the first end and has a homology arm with a length equal to or more than 50 nt at the second end.
Lu discloses a method of introducing a linear DNA donor construct comprising a single homology arm into the genome of Dietzia sp. DQ12-45-1b. After entering the bacterial cell, the linear donor construct is self-cyclized by non-homologous end joining (NHEJ) and then recombined with the targeted chromosomal gene by homologous recombination (HR). See, e.g., Abstract, and fig. 7. The length of the single homology arm is 410 bp. See, fig. 2.
Accordingly, Lu discloses a method comprising the manipulative actions instantly recited in claims 1 and 14.
Claims 1 and 14 further recite that, when double-strand breaks of a targeted genomic DNA occur, non-homologous recombination occurs at one of the 5’-end or 3’-end of the foreign DNA, the first end, and homologous recombination occurs at the other end, the second end. The limitation is repeated in the last two lines of claims 1 and 14, reciting that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end.
As instructed by MPEP 2111.04, the broadest reasonable interpretation of a method (or process) claim having contingent limitations requires only those steps that must be performed and does not include steps that are not required to be performed because the condition(s) precedent are not met. See Ex parte Schulhauser, Appeal 2013-007847 (PTAB April 28, 2016) ("[i]f the condition for performing a contingent step is not satisfied, the performance recited by the step need not be carried out in order for the claimed method to be performed ... [t]he Examiner did not need to present evidence of the obviousness of the [ ] method steps of claim 1 that are not required to be performed under a broadest reasonable interpretation of the claim").
In this case, the claims recite that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end contingent upon introducing double-strand breaks (DSBs) into the targeted genomic DNA. The claims do not positively recite a step (i.e., manipulative action) of introducing DSBs into the targeted genomic DNA, e.g., by introducing a targeted genomic DNA-cleaving enzyme, e.g., a CRISPR/Cas9 system. Therefore, since the cited reference, Lu, does not disclose a step of introducing DSBs into the targeted genomic, the process of non-homologous recombination occurring at the first end and homologous recombination occurring at the second end is not required to be performed to satisfy the conditions of the instantly claimed invention. For these reasons, the contingent limitations of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
The preamble of claim 1 recites a method for editing a genome in a cell, and the preamble of claim 14 recites a method for producing a cell preparation for treating severe combined immunodeficiency.
As instructed by MPEP 2111.02, if the body of a claim fully and intrinsically sets forth all of the limitations of the claimed invention, and the preamble merely states, for example, the purpose or intended use of the invention, rather than any distinct definition of any of the claimed invention’s limitations, then the preamble is not considered a limitation and is of no significance to claim construction. Shoes by Firebug LLC v. Stride Rite Children’s Grp., LLC, 962 F.3d 1362, 2020 USPQ2d 10701 (Fed. Cir. 2020) (The court found that the preamble in one patent’s claim is limiting but is not in a related patent); Pitney Bowes, Inc. v. Hewlett-Packard Co., 182 F.3d 1298, 1305, 51 USPQ2d 1161, 1165 (Fed. Cir. 1999). See also Rowe v. Dror, 112 F.3d 473, 478, 42 USPQ2d 1550, 1553 (Fed. Cir. 1997) ("where a patentee defines a structurally complete invention in the claim body and uses the preamble only to state a purpose or intended use for the invention, the preamble is not a claim limitation"); Kropa v. Robie, 187 F.2d at 152, 88 USPQ2d at 480-81 (preamble is not a limitation where claim is directed to a product and the preamble merely recites a property inherent in an old product defined by the remainder of the claim); STX LLC. v. Brine, 211 F.3d 588, 591, 54 USPQ2d 1347, 1350 (Fed. Cir. 2000) (holding that the preamble phrase "which provides improved playing and handling characteristics" in a claim drawn to a head for a lacrosse stick was not a claim limitation).
In this case, the preambles of claims 1 and 14 describe a purpose and/or intended use of performing the instantly claimed process steps (i.e., manipulative actions) recited in the body of the claims. The preambles do not positively recite any distinct definition of any of the claimed invention’s limitations, e.g., the nature of the genomic modification introduced into the host cell, if any, or the transgene encoded by the foreign DNA, if any, or the animal species or cell type of the genetically-modified cell, if any, or whether a modified cell is administered to a subject having severe combined immunodeficiency, etc. Moreover, Lu teaches that the DNA donor construct modifies the genome of a cell, wherein, after entering the bacterial cell, the linear donor construct is self-cyclized by non-homologous end joining (NHEJ) and then recombined with the targeted chromosomal gene by homologous recombination (HR). See, e.g., Abstract, and fig. 7. For these reasons, the purpose and/or intended use recited by the preambles of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
Accordingly, claims 1 and 14 are anticipated by the prior art.
Regarding dependent claims 6 and 15, Lu teaches the length of the single homology arm is 410 bp. See, fig. 2.
For these reasons, claims 1, 6, 14-15 are anticipated by the prior art. Dependent claims 3, 27-28 are not included in the basis of this rejection because the claims limit the cell to a blood cell, an undifferentiated cell, a differentiated cell, a nerve cell or a bone marrow cell, which is not taught or fairly suggested by the cited reference.
Claims 1, 3, 6, 14-15 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by US 2002/0116726 A1 to Croce et al.
This rejection is newly applied, necessitated by amendment.
Claims 1 and 14 are directed to methods comprising a step (i.e., manipulative action) of introducing foreign DNA comprising a 5’-end and a 3’-end into a targeted genome of a cell, wherein the foreign DNA does not have a homology arm at the first end and has a homology arm with a length equal to or more than 50 nt at the second end.
Croce discloses nonhuman transgenic animals with a disrupted FHIT gene. An endogenous FHIT locus in the genome of a mammalian cell is disrupted via homologous recombination by introducing a targeting DNA construct comprising at least one homology region. The nonhomologous portions (e.g., a transgene) may be flanked on each side by homology regions, or a single-flanking homology region may be used. See, Abstract, and par. 108. The targeting DNA construct is linear. See, par. 98, 113. The length of the homology regions is generally at least about 50 to 100 bases long, at least about 100 to 500 bases long, or at least about 750 to 2000 bases long. See, par. 98, 107.
Accordingly, Croce discloses a method comprising the manipulative actions instantly recited in claims 1 and 14.
Claims 1 and 14 further recite that, when double-strand breaks of a targeted genomic DNA occur, non-homologous recombination occurs at one of the 5’-end or 3’-end of the foreign DNA, the first end, and homologous recombination occurs at the other end, the second end. The limitation is repeated in the last two lines of claims 1 and 14, reciting that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end.
As instructed by MPEP 2111.04, the broadest reasonable interpretation of a method (or process) claim having contingent limitations requires only those steps that must be performed and does not include steps that are not required to be performed because the condition(s) precedent are not met. See Ex parte Schulhauser, Appeal 2013-007847 (PTAB April 28, 2016) ("[i]f the condition for performing a contingent step is not satisfied, the performance recited by the step need not be carried out in order for the claimed method to be performed ... [t]he Examiner did not need to present evidence of the obviousness of the [ ] method steps of claim 1 that are not required to be performed under a broadest reasonable interpretation of the claim").
In this case, the claims recite that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end contingent upon introducing double-strand breaks (DSBs) into the targeted genomic DNA. The claims do not positively recite a step (i.e., manipulative action) of introducing DSBs into the targeted genomic DNA, e.g., by introducing a targeted genomic DNA-cleaving enzyme, e.g., a CRISPR/Cas9 system. Therefore, since the cited reference, Croce, does not disclose a step of introducing DSBs into the targeted genomic, the process of non-homologous recombination occurring at the first end and homologous recombination occurring at the second end is not required to be performed to satisfy the conditions of the instantly claimed invention. For these reasons, the contingent limitations of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
The preamble of claim 1 recites a method for editing a genome in a cell, and the preamble of claim 14 recites a method for producing a cell preparation for treating severe combined immunodeficiency.
As instructed by MPEP 2111.02, if the body of a claim fully and intrinsically sets forth all of the limitations of the claimed invention, and the preamble merely states, for example, the purpose or intended use of the invention, rather than any distinct definition of any of the claimed invention’s limitations, then the preamble is not considered a limitation and is of no significance to claim construction. Shoes by Firebug LLC v. Stride Rite Children’s Grp., LLC, 962 F.3d 1362, 2020 USPQ2d 10701 (Fed. Cir. 2020) (The court found that the preamble in one patent’s claim is limiting but is not in a related patent); Pitney Bowes, Inc. v. Hewlett-Packard Co., 182 F.3d 1298, 1305, 51 USPQ2d 1161, 1165 (Fed. Cir. 1999). See also Rowe v. Dror, 112 F.3d 473, 478, 42 USPQ2d 1550, 1553 (Fed. Cir. 1997) ("where a patentee defines a structurally complete invention in the claim body and uses the preamble only to state a purpose or intended use for the invention, the preamble is not a claim limitation"); Kropa v. Robie, 187 F.2d at 152, 88 USPQ2d at 480-81 (preamble is not a limitation where claim is directed to a product and the preamble merely recites a property inherent in an old product defined by the remainder of the claim); STX LLC. v. Brine, 211 F.3d 588, 591, 54 USPQ2d 1347, 1350 (Fed. Cir. 2000) (holding that the preamble phrase "which provides improved playing and handling characteristics" in a claim drawn to a head for a lacrosse stick was not a claim limitation).
In this case, the preambles of claims 1 and 14 describe a purpose and/or intended use of performing the instantly claimed process steps (i.e., manipulative actions) recited in the body of the claims. The preambles do not positively recite any distinct definition of any of the claimed invention’s limitations, e.g., the nature of the genomic modification introduced into the host cell, if any, or the transgene encoded by the foreign DNA, if any, or the animal species or cell type of the genetically-modified cell, if any, or whether a modified cell is administered to a subject having severe combined immunodeficiency, etc. Moreover, Croce teaches that the DNA donor construct modifies the genome of a cell via homologous recombination. See, e.g., Abstract, and par. 36. For these reasons, the purpose and/or intended use recited by the preambles of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
Accordingly, claims 1 and 14 are anticipated by the prior art.
Regarding dependent claim 3, Croce teaches the cell is a pluripotent stem cell or an embryonic stem cell. See, par. 7, 55, 115.
Regarding dependent claims 6 and 15, Croce teaches the length of the homology arm is generally at least about 50 to 100 bases long, at least about 100 to 500 bases long, or at least about 750 to 2000 bases long. See, par. 98, 107.
For these reasons, claims 1, 3, 6, 14-15 are anticipated by the prior art. Dependent claims 27-28 are not included in the basis of this rejection because the claims limit the cell to a differentiated cell, a nerve cell or a bone marrow cell, which is not taught or fairly suggested by the cited reference.
Claims 1, 6, 14-15 and 27 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by US 2016/0152669 A1 to Nishi et al.
This rejection is newly applied, necessitated by amendment.
Claims 1 and 14 are directed to methods comprising a step (i.e., manipulative action) of introducing foreign DNA comprising a 5’-end and a 3’-end into a targeted genome of a cell, wherein the foreign DNA does not have a homology arm at the first end and has a homology arm with a length equal to or more than 50 nt at the second end.
Nishi discloses a means of producing a protein of interest from a transformed host cell by introducing a vector comprising a transgene into the host cell. The vector may be integrated into the host cell genome via non-specific integration without using homologous regions, integration using a single homologous region, or double-integration using two homologous regions. When integration uses a homologous region, the vector is linearized with restriction enzymes such that the homologous region is located at the end of the linear vector. See, Abstract; paragraphs 59-63. The length of the homologous region is 50 bp or longer. See, par. 62.
Accordingly, Nishi discloses a method comprising the manipulative actions instantly recited in claims 1 and 14.
Claims 1 and 14 further recite that, when double-strand breaks of a targeted genomic DNA occur, non-homologous recombination occurs at one of the 5’-end or 3’-end of the foreign DNA, the first end, and homologous recombination occurs at the other end, the second end. The limitation is repeated in the last two lines of claims 1 and 14, reciting that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end.
As instructed by MPEP 2111.04, the broadest reasonable interpretation of a method (or process) claim having contingent limitations requires only those steps that must be performed and does not include steps that are not required to be performed because the condition(s) precedent are not met. See Ex parte Schulhauser, Appeal 2013-007847 (PTAB April 28, 2016) ("[i]f the condition for performing a contingent step is not satisfied, the performance recited by the step need not be carried out in order for the claimed method to be performed ... [t]he Examiner did not need to present evidence of the obviousness of the [ ] method steps of claim 1 that are not required to be performed under a broadest reasonable interpretation of the claim").
In this case, the claims recite that non-homologous recombination occurs at the first end and homologous recombination occurs at the second end contingent upon introducing double-strand breaks (DSBs) into the targeted genomic DNA. The claims do not positively recite a step (i.e., manipulative action) of introducing DSBs into the targeted genomic DNA, e.g., by introducing a targeted genomic DNA-cleaving enzyme, e.g., a CRISPR/Cas9 system. Therefore, since the cited reference, Nishi, does not disclose a step of introducing DSBs into the targeted genomic, the process of non-homologous recombination occurring at the first end and homologous recombination occurring at the second end is not required to be performed to satisfy the conditions of the instantly claimed invention. For these reasons, the contingent limitations of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
The preamble of claim 1 recites a method for editing a genome in a cell, and the preamble of claim 14 recites a method for producing a cell preparation for treating severe combined immunodeficiency.
As instructed by MPEP 2111.02, if the body of a claim fully and intrinsically sets forth all of the limitations of the claimed invention, and the preamble merely states, for example, the purpose or intended use of the invention, rather than any distinct definition of any of the claimed invention’s limitations, then the preamble is not considered a limitation and is of no significance to claim construction. Shoes by Firebug LLC v. Stride Rite Children’s Grp., LLC, 962 F.3d 1362, 2020 USPQ2d 10701 (Fed. Cir. 2020) (The court found that the preamble in one patent’s claim is limiting but is not in a related patent); Pitney Bowes, Inc. v. Hewlett-Packard Co., 182 F.3d 1298, 1305, 51 USPQ2d 1161, 1165 (Fed. Cir. 1999). See also Rowe v. Dror, 112 F.3d 473, 478, 42 USPQ2d 1550, 1553 (Fed. Cir. 1997) ("where a patentee defines a structurally complete invention in the claim body and uses the preamble only to state a purpose or intended use for the invention, the preamble is not a claim limitation"); Kropa v. Robie, 187 F.2d at 152, 88 USPQ2d at 480-81 (preamble is not a limitation where claim is directed to a product and the preamble merely recites a property inherent in an old product defined by the remainder of the claim); STX LLC. v. Brine, 211 F.3d 588, 591, 54 USPQ2d 1347, 1350 (Fed. Cir. 2000) (holding that the preamble phrase "which provides improved playing and handling characteristics" in a claim drawn to a head for a lacrosse stick was not a claim limitation).
In this case, the preambles of claims 1 and 14 describe a purpose and/or intended use of performing the instantly claimed process steps (i.e., manipulative actions) recited in the body of the claims. The preambles do not positively recite any distinct definition of any of the claimed invention’s limitations, e.g., the nature of the genomic modification introduced into the host cell, if any, or the transgene encoded by the foreign DNA, if any, or the animal species or cell type of the genetically-modified cell, if any, or whether a modified cell is administered to a subject having severe combined immunodeficiency, etc. Moreover, Nishi teaches that the DNA donor construct modifies the genome of a cell via homologous recombination. See, e.g., Abstract, and par. 29. For these reasons, the purpose and/or intended use recited by the preambles of claims 1 and 14 are not found to patentably distinguish the instantly claimed invention from the prior art.
Accordingly, claims 1 and 14 are anticipated by the prior art.
Regarding dependent claims 6 and 15, Nishi teaches the length of the homologous region is 50 bp or longer. See, par. 62.
Regarding dependent claim 27, Nishi teaches the cell is an animal cell or a Chinese hamster ovary (CHO) cell. See, par. 2, 51.
For these reasons, claims 1, 6, 14-15 and 27 are anticipated by the prior art. Dependent claims 3 and 28 are not included in the basis of this rejection because the claims limit the cell to a blood cell, an undifferentiated cell, a nerve cell or a bone marrow cell, which is not taught or fairly suggested by the cited reference.
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.
Claims 3 and 28 are rejected under 35 U.S.C. 103 as being unpatentable US 2016/0152669 A1 to Nishi et al., as applied to claims 1, 6, 14-15 and 27 above; in further view of US 2012/0208233 A1 to Schweiger et al.
Nishi teaches the host cell is an animal cell or a Chinese hamster ovary (CHO) cell. See, par. 2, 51. Nishi does not teach or fairly suggest that the host cell is a blood cell, an undifferentiated cell, a nerve cell or a bone marrow cell, as claimed in claims 3 and 28.
Schweiger is relevant prior art for disclosing methods of genetically modifying host cells by introducing an exogenous DNA donor construct for the expression of proteins. See, e.g., Abstract, and par. 1-8. Suitable mammalian host cells include blood cells (e.g., Lymphoid cell lines, leukemia cell lines, primary CD34+ haematopoetic cell lines), undifferentiated cells (e.g., Embryonic kidney cell lines, stem cell lines, embryonic fibroblast cell lines), nerve cells (e.g., Neuroblastoma cell lines, rat DRG neuron cell lines) and bone marrow cells (e.g., Osteosarcoma cell lines, lymphoid cell lines, primary CD34+ haematopoetic cell lines). See, par 46, and Table 1 on pg. 4.
Accordingly, prior to the effective filing date of the instantly claimed invention, one of ordinary skill in the art would have recognized that blood cells, undifferentiated cells, nerve cells and bone marrow cells were suitable alternative host cells for recombinant protein production. Therefore, it would have been prima facie obvious to one of ordinary skill in the art to substitute animal cells or CHO cells, as taught by Nishi, with blood cells, undifferentiated cells, nerve cells or bone marrow cells, as taught by Schweiger, with a reasonable expectation of success because said cell lines provide suitable expression systems for recombinant protein production, and the simple substitution of one known element for another would have yielded predictable results to one of ordinary skill in the art at the time of the invention.
Conclusion
The following publications made of record and not relied upon are considered pertinent to applicant's disclosure:
Suzuki et al. (2016) “In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration” Nature, 540(7631), 144-149, discloses a homology-independent targeted integration (HITI) strategy which allows for robust DNA knock-in in both dividing and non-dividing cells in vitro and, more importantly, in vivo (for example, in neurons of postnatal mammals). The donor templates comprise HITI sequences on both homology arms, and the homology arms are asymmetrical in length. See, e.g., Abstract, and fig. 1a.
US 2020/0354734 A1 to Niessen et al. claims priority to 10/30/2017 based on U.S. Provisional Application No. 62/578,621. The disclosure provides methods for precision genome editing (GE) in eukaryotic cells by inactivating the non-homologous end-joining (NHEJ) DNA repair pathway. A targeting vector is introduced into the genome of a host cell, wherein the targeting vector may comprise only one homology region. A vector using a single homology region will usually have a higher degree of complementarity compared to a vector using more than one homology region in order to allow hybridization. See, Abstract, and paragraph 96; see also page 29 of the provisional application.
US 5,698,763 A to Weissman et al., issued on 12/16/1997, discloses means of making transgenic mammals and birds by introducing DNA targeting molecules that specifically disrupt endogenous prion protein (PrP) genes by homologous recombination into animal cells. The targeting molecule is a linear DNA molecule or a circular DNA molecule. A circular targeting molecule could comprise a pair of homologous regions separated by a disrupting region. Alternatively, a circular targeting molecule could comprise a single homologous region. Upon integration at the target gene locus, the circular molecule would become linearized, with a portion of the homologous region at each end. Thus, the single homologous region effectively becomes two homologous regions, as in the case of linear targeting molecules See, Abstract, and col. 6, ll. 47-67.
US 2022/0333106 A1 to Izpisua Belmonte et al., filed after the effective filing date of the instantly claimed invention, describes methods for editing a target genome in a cell by contacting the cell with (i) a single homology arm construct comprising a replacement sequence and a targeted endonuclease cleavage site; and (ii) a targeted endonuclease, wherein the replacement sequence comprises at least one nucleotide difference compared to the target genome and wherein the target genome comprises a sequence homologous to the targeted endonuclease cleavage site. In particular, the disclosure provides a unique non-homologous end joining (NHEJ) and homology-directed repair (HDR) mediated targeted gene knock-in method that requires a double-strand break (DSB) induction site within a single stretch of homologous sequence on the donor. This design is termed “intercellular linearized Single homology Arm donor mediated intron-Targeting Integration (SATI)”. SATI allows DNA knock-in via single homology arm mediated HDR or homology independent NHEJ-based HITI, enabling targeting a broad range of mutations and cell types. FIG. 6B shows a schematic representation of the new intronic gene-knock-in method, SATI. The donor DNA includes DSB induction site and one-homology arm where is identical to the target genome. SATI allows DNA knock-in via single homology arm mediated HDR (oaHDR) or homology independent NHEJ-based HITI, enabling to target a broad range of mutations and cell types. See also, Figures 1A, 2A, and 6B.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAMES J GRABER whose telephone number is (571)270-3988. The examiner can normally be reached Monday-Thursday: 9:00 am - 4:00 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, James D Schultz can be reached on (571)272-0763. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/JAMES JOSEPH GRABER/Examiner, Art Unit 1631
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