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 affidavit of 4/13/2026 by DE VLEESSCHAUWER is acknowledged and has been fully considered.
The applicant’s response filed 4/13/2026 has been entered.
Claims 1-2, 6-7, 10-12, 17-18, 21-23, 25 have been amended.
New claims 26-30 have been added.
Notes: On 5/6/2024, the applicant elected species:
Election 1: RNA guided nuclease,
Election 1a: Cas12a and Cas9.
Election 2: Single guide RNA (sgRNA). Note by the examiner on 6/24/2026: the newly added guide RNA (gRNA) is also examined.
In summary, claims 1-30 are pending and examined in the office action. Non-elected species are withdrawn.
All previous objections and rejections not set forth below have been withdrawn in view of the applicant’s amendment and/or upon further consideration. See “Response to Arguments” at the end of office action.
The following rejections are repeated, modified and/or added for the reasons of record as set forth in the last Office action of 11/12/2025, and/or necessitated by the applicant’s amendments. The applicant’s arguments filed 4/13/2026 have been thoroughly considered but are not deemed fully persuasive.
Claim Rejections - 35 USC § 112
Indefiniteness
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.
Amended claims 1-2 and dependent claims 3-30 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 pre-AIA the applicant regards as the invention.
Both claims 1 and 2 are amended to recite “wherein said donor DNA molecule comprises a DNA fragment of the target region and contains the donor DNA that differs from the wild-type sequence of the target region”.
The claims are contradicting to the specification and the art, because:
The specification (page 13, 3rd para) defines donor DNA molecule: As used herein the terms "donor DNA molecule", "repair DNA molecule" or "template DNA molecule" all used interchangeably herein mean a DNA molecule having a sequence that is to be introduced into the genome of a cell. It may be flanked at the 5' and/or 3' end by sequences homologous or identical to sequences in the target region of the genome of said cell.
Accordingly, according to the specification and the general understanding of the art, a “donor DNA molecule” to be introduced to a native genome; before the “donor DNA molecule” has been introduced and integrated into the native genome, it does not (and should not) comprise a DNA fragment of the target region or any region of the genome.
Therefore, the term recited in the claim is contradicting to the specification and to the general understanding of the art.
Appropriate corrections and clarifications are required for one skill in the art to understand the claim to carry out the invention.
Interpretation of “donor DNA molecule” and “donor DNA” in claims
According to the specification (page 13, 3rd para) the terms "donor DNA molecule" means a DNA molecule having a sequence that is to be introduced into the genome of a cell. It may be flanked at the 5' and/or 3' end by sequences homologous or identical to sequences in the target region of the genome of said cell.
Note: identical to a sequence in the target region is not same as comprise a DNA fragment of the target region.
As analyzed above, “wherein said donor DNA molecule comprises a DNA fragment of the target region” is deemed indefinite.
For compact prosecution and definiteness, in view of the specification, the “said donor DNA molecule comprises a DNA fragment of the target region” is interpreted as --- said donor DNA molecule comprises a DNA fragment identical to a sequence of the target region ---.
The first wherein clause in each of claim 1 and claim 2 is interpreted as
--- wherein said donor DNA molecule
(1) comprises a DNA fragment identical to a sequence of the target region, and
(2) contains/comprises the donor DNA that differs from the wild-type sequence of the target region flanked by at least 30 bases that are each at least 80% identical to the sequence in the target region ---.
Note: from the context of claims, a “donor DNA molecule” is bigger than a “donor DNA”, and comprises the “donor DNA”.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries set forth in Graham v. John Deere Co., 383 U.S. 1, 148 USPQ 459 (1966), that are applied 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 non-obviousness.
Claims 1-7, 11-18, 22-24 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al (Targeted mutagenesis using the Agrobacterium tumefaciens-mediated CRISPR-Cas9 system in common wheat. BMC Plant Biology, 1-12, 2018), in view of Arndell et al (Research Article. gRNA validation for wheat genome editing with the CRISPR-Cas9 system. BMC Biotechnology, p1-12, 2019.10), and Yamamoto et al (Making ends meet: Targeted integration of DNA fragments by genome editing. Chromosoma. 127(4): 405–420, 2018).
The PDF of the publish date of Arndell et al is also attached.
Amended claim 1 is drawn to a method comprising the steps of
a. introducing into a wheat cell
i. at least one donor DNA molecule, and
ii. at least one RNA guided nuclease or RNA guided nickase, and
iii. an RNA of: at least one singleguideRNA (sgRNA) or guideRNA (gRNA); or at least trans-activating CRISPR RNA (tracrRNA) and CRISPR RNA (crRNA), and
b. incubating the wheat cell to allow for introduction of said at least one donor DNAinto said target region of the genome by homologous recombination, and
c. selecting a wheat cell comprising said precise edit in said target region, said precise edit comprising the sequence of the donor DNA in said target region,
wherein said donor DNA molecule
(1) comprises a DNA fragment identical to a sequence of the target region, and
(2) contains/comprises the donor DNA that differs from the wild-type sequence of the target region flanked by at least 30 bases that are each at least 80% identical to the sequence in the target region,
wherein said donor DNA molecule is used as repair template for homologous recombination to introduce the donor DNA into the target region, and
wherein said precise edit consists of the donor DNA comprises said precise edit, and is being introduced in the target region without any InDels, duplications or other mutations as compared to the unaltered DNA sequence of the target region that are not comprised in the donor DNA molecule sequence.
for making a precise edit in a target region of the genome of wheat through introduction of at least one donor DNA into said target region (preamble).
Amended claim 2 is drawn to a method comprising the steps of
a. introducing into a wheat cell
i. at least one donor DNA molecule, and
ii. at least one RNA guided nuclease or RNA guided nickase, and
iii. an RNA of: at least one singleguideRNA (sgRNA) or guideRNA (gRNA); or at least trans-activating CRISPR RNA (tracrRNA) and CRISPR RNA (crRNA), and
b. incubating the wheat cell to allow for introduction of said at least one donor DNA into said target region of the genome by homologous recombination,
c. selecting a wheat cell comprising said precise edit in said target region, and
d. regenerating a wheat plant from said selected wheat cell,
wherein said donor DNA molecule
(1) comprises a DNA fragment identical to a sequence of the target region, and
(2) contains/comprises the donor DNA that differs from the wild-type sequence of the target region flanked by at least 30 bases that are each at least 80% identical to the sequence in the target region,
wherein said donor DNA molecule is used as repair template for homologous recombination to introduce the donor DNA into the target region, and
wherein said precise edit consists of the donor DNA comprises said precise edit, and is being introduced in the target region without any InDels, duplications or other mutations as compared to the unaltered DNA sequence of the target region that are not comprised in the donor DNA molecule sequence,
for producing a wheat plant comprising a precise edit in a target region of its genome though introduction of donor DNA in said target region (preamble).
Notes:
A. Claims are somewhat broader after the amendment: sgRNA is no longer solely required, any guide RNA (gRNA) is fine.
B. Claim 2 is fundamentally the same as claim 1, except:
1) step c does not require “said precise edit comprising the sequence of the donor DNA in said target region”, and
2) requires step d. d. regenerating a wheat plant from said selected wheat cell.
Zhang et al teach a method of introduction of donor DNA molecules into a target region of the genome of wheat, and producing a wheat plant comprising a donor DNA (inserted/introduced DNA) in the target region by using a CRISPR/Cas9 system. “CRISPR/Cas9 system has been widely used to precisely edit plant genomes”. Thus, the edit is considered precise (p1, Abstract).
Specifically, the method comprises introducing into a wheat cell/protoplast target genes for insertion or deletion/donor DNA molecule;
a gene encoding and expressing Cas9 (reading on the limitations of claims 1-2, 4-5, 15-16); and
sgRNAs (synthetic guide RNA, p7, fig 3)
(p1, abstract; p2, right col, Methods).
The wheat cells are incubated with the target genes/donor DNAs to allow the introduction of the target genes (p3, left col, 1st para).
The cells having the mutations (targeted insertions or deletions) are selected (by PCR, p3, left col, 2nd and 3rd paras, right col, 1st para). “CRISPR/Cas9 system has been widely used to precisely edit plant genomes” (p1, Abstract). Thus, the edit is considered precise.
A plasmid comprises an expression cassette comprising the genes/donor DNAs, the Cas9 encoding DNA sequence and the sgRNA encoding sequence is used for the introduction (p2, right col, 2nd para).
The target genes/donor DNAs are pre-selected for efficient targeting (p2, left col, 3rd para); and the sgRNAs are also pre-selected and designed to recognize the specific region of the coding sequence of the target genes for efficient targeting (p3, right col, last para, p4, left col, 1st para).
The site-specific introduction was successful, and the target genes are in the wheat genome, the efficiency was as high as 54.17% (Results in p3, right col, last para; p4-5, whole pages; p10, right col, last para).
Zhang et al further suggest that the method can be used for inserting transgenes (reading on donor DNA molecule, p7, left col, 1st para).
Zhang et al teach and demonstrated that the lines comprising the targeted mutants and transgenes were successfully regenerated (p5, left col, 1st para), teaching the step d. of claim 2. Thus, the producing of a wheat plant comprising the genes/donors in the genome was also successful.
Zhang et al further teach and demonstrated that “off-target mutations were not detected in regions that were highly homologous to the sgRNA sequences” (p1, Abstract; p5, right col, 2nd para; p10, left col, 1st para). Thus, Zhang et al teach “the target region without any indels or other mutations”. Also, the process of Zhang et al at least comprise some part of homologous recombination. Zhang et al teach a strong motivation to perform homologous recombination to avoid or reduce off-target indels and mutations, and teach an advantage of homologous recombination.
Thus, Zhang et al teach and/or suggest the claim limitations, except do not explicitly teach (but suggest nevertheless)
(1) the introduction of donor DNA is by homologous recombination; and
(2) said donor DNA is flanked by at least 30 bases that are each at least 80% identical to the sequence in the target region.
Arndell et al teach and demonstrated using a CRISPR-Cas9 system comprising guide RNA (gRNA) to edit wheat genome in a site-specific manner (p1, Abstract).
Specifically, Arndell et al teach and demonstrated using homologous recombination (HDR) to edit wheat genome (p2, left col, last para, whole right col; p7-8, Methods).
Arndell et al demonstrated mutations including insertion mutations in the target region of the wheat genome (p4, fig 2), reading on introducing donor DNA.
Arndell et al demonstrated no off-target mutations by the HDR in some of the vectors (p6, left col, 2nd para), reading on no unwanted indels, duplication and mutations.
Arndell et al further teach that non-homologous method is error-prone, while homologous directed repair (HDR) is precise (p1, right col). Hence, Arndell et al provide strong motivation to perform what Zhang et al suggested, and provide high expectation of success by using homologous recombination for wheat genome editing.
After instant claim amendment, sgRNA is no longer a claim requirement but an option. Arndell et al nevertheless also made and used sgRNA (p7, right col, last para).
Arndell et al are silent regarding the arm length.
Yamamoto et al teach using Crispr-Cas9, sgRNA and donor DNA to editing genomes including plant genomes (p2233, Abstract; p2, 1st para; p4, 3rd para). Yamamoto et al teach methods of homologous recombination (p9, 1st para to p12, 3rd para).
Yamamoto et al particularly teach a method of homologous recombination with short homology arms, in which the arm length is 30-100 bases homology (reading on at least 30 bases) with the target DNA, and demonstrated that homologous recombination (HDR) can work with only 50 bases of homology. Yamamoto et al also teach that the advantage is to facilitate the introduction of the donor DNA, and increase the homology-directed repair (HDR) (p10, 2nd para to p11, 2nd para). Note: any inserted sequence is a donor DNA sequence. Since both the donor DNA sequence and the target region of the wheat genome comprise the at least 30 base sequence that is homologous (or at least 80% identical), the target region and the donor DNA are immediately/directly flanking to each other.
Regarding dependent claims, Arndell et al teach including a selectable glyphosate resistance as a robust selectable marker in wheat tissue culture and during plant growth (p5, right col, 1st para), the limitation of claims 3 and 14.
Zhang et al teach a plasmid comprising an expression cassette comprising genes/donor DNAs, Cas9 encoding DNA sequence, and the sgRNA encoding sequence, is used for the introduction (p2, right col, 2nd para), reading on the limitation of claims 6-7 and 17-18.
Zhang et al teach that the target genes/donor DNAs are pre-selected for efficient targeting (p2, left col, 3rd para); and the sgRNAs are also pre-selected and designed to recognize the specific region of the coding sequence of the target genes for efficient targeting (p3, right col, last para, p4, left col, 1st para), reading on the limitations of claims 11 and 22.
Zhang et al teach that Agrobacterium carrying the expression cassette is used for introduction/transformation (p3, left col, 1st para), reading on the limitations of claims 12 and 23.
Arndell et al teach and demonstrated including a nuclear localization signal in the vector (p8, left col, 2nd para), the limitation of claims 13 and 24.
An invention would have been obvious to one ordinary skill in the art if any teaching, suggestion or motivation in prior art leading the one to combine the teaching(s) or suggestion(s) of the cited references to arrive the claimed invention.
In this case, it would have been obvious for one ordinary skill in the art to modify the invention of Zhang et al, such that the method is/or include editing by homologous recombination as suggested by Zhang et al and taught and demonstrated by Arndell et al in wheat and Yamamoto et al in plants, and the donor DNA comprises an arm of at least 30 bases identical to a sequence in the target region also as taught by Yamamoto et al. One ordinary skill in the art would have been motivated to do so for the advantage of reducing off-target mutations (up to no off-site mutation) as taught and demonstrated by Arndell et al in wheat. The expectation of success would have been high because Arndell et al demonstrated the success of donor insertion with low to none off-target mutations.
Therefore, the invention would have been obvious to one ordinary skill in the art.
Claims 8 and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al in view of Arndell et al and Yamamoto et al, as applied to claims 1-2, 6 and 17 above, and further in view of HX Zhang et al (Genome Editing with mRNA Encoding ZFN, TALEN, and Cas9. Molecular Therapy Vol. 27, p735-746, 4/2019).
Claims 1-2, 6 and 17 have been analyzed above. Claims 6 and 17 recite that the at least one nuclease or the at least one single guide RNA (sgRNA, elected species) is introduced into said cell encoded by a nucleic acid molecule.
Claims 8 and 19 depend on claims 6 and 17, wherein the nucleic acid is an RNA molecule.
Zhang et al teach that the nuclease and sgRNA are encoded by a nucleic acid molecule, but Zhang et al in view of Yamamoto et al, Filippova et al, and Upadhyay et al do not teach “by an RNA molecule”.
HX Zhang et al teach that Cas9 is encoded by DNA in cell including plant cells (p735-p736, Principles of Genomic Editing).
HX Zhang et al also teach that Cas9 is encoded by mRNA in vitro (p735, Abstract; p737, right col, last para; p738, left col, 1st para). HX Zhang et al further teach and demonstrated that the in vitro mRNA encoded nucleases had produced high editing efficiency (p740, Table 3).
It would have been obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al and Yamamoto et al, such that the nuclease/Cas are encoded by RNA molecules in vitro as an alternative of by DNA molecules in plant cells, as taught by HX Zhang et al. One ordinary skill in the art would have been motivated to do so because HX Zhang et al demonstrated that the mRNA encoded nucleases had produced high editing efficiency in vitro. The expectation of success would have been high because the in vitro mRNA encoded nucleases had been demonstrated and had produced high efficiency of editing by HX Zhang et al.
In addition, according to the specification (p10, 5th para), an RNA molecule may be merely an alternative to a DNA molecule for encoding a nuclease or a sgRNA. Thus, the applicant merely claims what the prior art had taught.
Therefore, the claims would have been obvious to one ordinary skill in the art.
Claims 9 and 20 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al in view of Arndell et al and Yamamoto et al as applied to claims 1-2, 6 and 17 above, and further in view of Gao et al (WO 2018202199, published 11/8/2018, filed 5/7/2018).
Claims 1-2, 6 and 17 have been analyzed above.
Claims 9 and 20 depend on claims 6 and 17, wherein a polynucleotide sequence encoding the at least one nuclease is sequence optimized for expression in wheat.
Zhang et al in view of Arndell et al and Yamamoto et al do not teach codon optimization for wheat expression.
Gao et al teach a method comprising using Cas9 to edit plants including wheat (Example 7 in p69, last para). Gao et al teach codon optimization and teach the optimized sequence of the DNA of interest for expression in cereal plants (p17, SEQ ID NO: 2; p18, SEQ ID NO: 14).
Gao et al teach the advantage that "Codon optimization" implies that the codon usage of a DNA or RNA is adapted to that of a cell or organism of interest to improve the transcription rate of said recombinant nucleic acid in the cell or organism of interest. The skilled person is well aware of the fact that a target nucleic acid can be modified at one position due to the codon degeneracy, whereas this modification will still lead to the same amino acid sequence at that position after translation, which is achieved by codon optimization to take into consideration the species-specific codon usage of a target cell or organism” (p29, 1st para).
It would have been prima facie obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al and Yamamoto et al, such that the coding sequence of the DNA of interest is optimized for the expression in the specific organism, as taught by Gao et al. One ordinary skill in the art would have been motivated to do so because the codon optimization improves the transcription rate of said recombinant nucleic acid in the cell or organism of interest, as taught by Gao et al. The expectation of success would have been high because codon optimization had been a mature method as taught by Gao et al, for example.
Therefore, the claims would have been obvious to one ordinary skill in the art.
Claims 10 and 21 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al in view of Arndell et al and Yamamoto et al, as applied to claims 1-2 above, and further in view of Liang et al (Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nature Communication. p1-5, 2016).
Claims 1-2 have been analyzed above.
Claims 10 and 21: wherein the at least one RNA guided nuclease and the at least one sgRNA are introduced into said cell as ribonucleoprotein (RNP) assembled outside said cell (Note: no donor DNA is in the RNP).
Zhang et al additionally suggest by citing references that Cas9 and guide RNA can be formed to a ribonucleoprotein complex (p10, left col, last para). Arndell et al also suggest using Cas9 ribonucleoprotein in the system (p7, left col, 1st para).
Thus, Zhang et al in view of Arndell et al and Yamamoto et al suggest but do not explicitly teach such a ribonucleoprotein complex.
Liang et al teach making a Cas9/sgRNA ribonucleoprotein (RNP) complex before the RNP is introduced to wheat cells by bombardment (p4, right col, last para; p5, left col, 1st para).
Liang et al further teach that “the most important advantage of CRISPR/Cas9 RNP mediated genome editing is the elimination of transgene integration and small DNA insertions in the mutants generated (p4, left col, 3rd para).
Liang et al achieved success of the DNA-free editing (p1, Abstract; p2-4, Results).
It would have been prima facie obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al and Yamamoto et al, such that the nuclease and the sgRNA is formed to a RNP outside of the cells, and the RNP is delivered to the wheat cells, as suggested by Zhang et al and Arndell et al and taught by Liang et al. One ordinary skill in the art would have been motivated to do so to take the advantage of the elimination of transgene integration and small DNA insertions in the mutants generated as taught by Liang et al. The expectation of success would have been high because such RNP had been made and delivered to wheat cells, and achieved successful DNA-free editing, as demonstrated by Liang et al.
Therefore, the claims would have been obvious to one ordinary skill in the art.
New claims 26-29 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al in view of Arndell et al and Yamamoto et al, as applied to claims 2, and further in view of Labs (WO 2018138385, published 8/2/2018, filed 1/30/2018).
New claims 26-27 limit claim 2, wherein said precise edit in said donor DNA molecule is flanked by at least 150 bases that are each at least 80% identical and 100% to the sequence in the target gene.
New Claims 28 limits claim 26, wherein said donor DNA comprises silent mutations to prevent cleavage of the donor DNA and the edited allele with the desired precise edit.
New Claim 29 limits claim 26, wherein said precise edit in said donor DNA molecule is flanked by at least 350 bases that are each identical to the sequence in the target gene and contains some silent mutations to prevent cleavage of the donor DNA and the edited allele with the desired edit.
Zhang et al in view of Arndell et al and Yamamoto et al do not teach such limitations.
Labs teaches a method of using CRISPR and guide RNA to edit genomes (p2, 2nd para) including plants and wheat (p4, end para).
Labs teaches that the arm/flanking region of the donor molecule can comprise 150, 500, 1000, and up to 5000 bases homologous to the sequence of a target region. Labs teaches the advantage thereof being for the optimized recognition (p62, 1st para). Thus, Labs at least suggests the limitation, and teach the advantage thereof, teaching claims 26-27.
Labs teaches including a silent mutation to prevent and not affect the donor DNA’s encoding sequence (p52, 2nd para), teaching claim 28.
The at least 500 bases above reads on the limitation of claim 29.
It would have been prima facie obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al and Yamamoto et al, such that the flanking sequence is at least 150 bases long, or at least 350 bases long, as suggested by Labs. One ordinary skill in the art would have been motivated to do so to take the same advantage of the optimized recognition as taught by Labs, to achieve the expected result, with a reasonable expectation of success.
It also would have been prima facie obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al and Yamamoto et al, such that the vector includes a silent mutation, as taught by Labs. One ordinary skill in the art would have been motivated to do so to take the same advantage of preventing cleavage of the donor DNA’s encoding sequence as taught by Labs, to achieve the expected result, with a reasonable expectation of success.
Therefore, the claims would have been obvious to one ordinary skill in the art.
Amended claim 25 and new claim 30 are rejected under 35 U.S.C. 103 as being unpatentable over Zhang et al in view of Arndell et al, Yamamoto et al, and Labs, as applied to claims 2 and 26, and further in view of Cui et al (An optimised CRISPR/Cas9 protocol to create targeted mutations in homoeologous genes and an efficient genotyping protocol to identify edited events in wheat. BMC Plant Methods. p1-12, 2019.10).
Claim 2 has been analyzed above.
Claim 25 limits claim 2, wherein said wheat comprises three subgenomes, wherein said target region comprises a target gene that has at least three homeologous copies in the wheat genome, and wherein one or two or three homeologous copies more alleles of said target gene in one or two or three moreof said subgenomes comprise said precise edit.
New claim 30 limits claim 26, wherein said sgRNA or gRNA (or tracrRNA and crRNA) target all homeologous copies of said target gene copies in all subgenomes, and wherein said donor DNA molecule comprises a DNA fragment of the target gene of one of the subgenomes and contains the donor DNA that differs from the wild-type sequence of the target region.
Zhang et al in view of Arndell et al, Yamamoto et al and Labs teach claim 2 but do not teach the subject matter of claims 25 and 30.
Cui et al teach a method of using CRISPR-Cas9 and sgRNA to edit homoeologous genes of wheat (p1, Abstract).
Cui et al teach that wheat genome has 3 subgenomes (A, B and D) (p2, left col, 2nd para), and teach using the CRISPR-Cas9 and sgRNA to edit homoeologous genes in the 3 subgenomes, and achieved success (p2, left col, 3rd to 4th para; p8-12, Methods; p2, right col, 1st para to p5, right col, 1st para).
Specifically, Cui et al teach a detail method of identifying edit events of three homoeologous genes in the three subgenomes (p6, fig 2). Cui et al demonstrated different editing efficiency, some genes were edited with high efficiencies (p3, right col, 2nd para to p4, left col, 1st para).
Notes: all three subgenomes are demonstrated to be edited.
Thus, it would have been obvious to one ordinary skill in the art to modify the invention rendered obvious by Zhang et al in view of Arndell et al, Yamamoto et al and Labs, such that the CRISPR-Cas and sgRNA system is used to target and edit all homeologous copies in the three wheat genomes as taught by Cui et al. One ordinary skill in the art would have been motivated to do so because Cui et al demonstrated success of different efficiencies in wheat. The expectation of success would have been high because Cui et al demonstrated success of all subgenomes being edited with different efficiencies in wheat. Thus, such modification would have been expected to be successful.
Therefore, the claims would have been obvious to one ordinary skill in the art.
Response to Arguments
The applicant significantly amended independent claims 1-2, and added new claims 26-30. The amended claims 1-2 are indefinite, as analyzed above.
The amended claims 1-2 are also broader for including guide RNA (gRNA) as an option, thus sgRNA is no longer required but is also an option.
The applicant’s arguments and the affidavit of 4/13/2026 by DE VLEESSCHAUWER is acknowledged and has been fully analyzed and considered. However, the arguments and the declaration are arguing that the previous cited reference(s) does/do not explicitly demonstrate using homologous recombination to target wheat plant, and that a low frequency of HDR remains a major challenge in plant genome editing in wheat. The affidavit cites multiple references indicating that homologous recombination to target wheat plant is still a challenge.
In view of such significant amendments, the rejections are significantly modified and/or re-written, citing new references, as analyzed above.
The arguments to the previous references are no longer applicable. The arguments are not persuasive in view of new rejections using new references. For compact prosecution, the examiner would bring the following critical point:
In brief, Arndell et al also recognize that “the inherent low frequency of HDR remains a major challenge in plant genome editing” (p2, left col, 1st para).
Arndell et al nevertheless teach and demonstrated using a CRISPR-Cas9 system comprising guide RNA (gRNA) to edit wheat genome in a site-specific manner (p1, Abstract). Specifically, Arndell et al teach and demonstrated using homologous recombination (HDR) to edit wheat genome (p2, left col, last para, whole right col; p7-8, Methods). Arndell et al demonstrated no off-target mutations by the HDR in some of the vectors (p6, left col, 2nd para), reading on no unwanted indels, duplication and mutations.
Hence, low frequency of HDR should not be a factor to prevent one ordinary skill in the art to use it, because HDR has the advantage of low off-target mutations, as taught by the cited references including Zhang et al and Arndell et al.
As a fact, Arndell et al recognized that but nevertheless used it. Any ordinary skill in the art would have done the same.
Moreover and critically, editing efficiency is not a claim requirement. Any slight amount of editing is deemed a success.
Conclusion
No claim is allowed.
The 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). The 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 extension fee 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 date of this final action.
Contact information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to WAYNE ZHONG whose telephone number is (571)270-0311. The examiner can normally be reached 8:30am to 5:00pm EST.
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/Wayne Zhong/
Primary Examiner, Art Unit 1662
Prosecution Insights