DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 04/07/2026 has been entered.
Claim Status
Claims 1-2, 4, 7-8, and 10 are pending.
Claims 7-8 are withdrawn from examination as being part of a nonelected group.
Claims 1-2, 4 and 10 are being examined.
All previous objections and rejections not set forth below have been withdrawn in view of applicant’s amendments to the claims.
Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Certified copy of the English translation of the foreign priority document has been received by the Office on 04/07/2026.
Claim Rejections - 35 USC § 103
Claims 1-2, 4 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over
Jansing et al. (CRISPR/Cas9-mediated knockout of six glycosyltransferase genes in Nicotiana benthamiana for the production of recombinant proteins lacking β-1,2-xylose and core α-1,3-fucose. 2019. Plant Biotechnology Journal, 17:350–361) and Steinkellner et al. (US 2020/0080100 A1), and in view of Kikuchi et al. (Collection, mapping, and annotation of over 28,000 cDNA clones from japonica rice, Science, 2003, 301:376-9), for reasons of record stated on pages 4-9 in the Office action dated 03/21/2025.
Claims 1-2 are drawn to a vector for editing rice N-glycosylation gene(s) comprising a polycistronic guide RNA targeting different endogenous genes encoding β1,2-xylosyltransferase (β1,2-XylT); β 1,3-galactosyltransferase (β1,3-GalT); α1,3-fucosyltransferase (αl,3-FucT); α1,4-fucosyltransferase (αl,4-FucT); and hexosaminidase (1-4) proteins.
Jansing et al. teaches an easily scalable alternative platform for the production of Pharmaceutical or therapeutic proteins by using a multiplex CRISPR/Cas9 based genome editing to generate tobacco (Nicotiana benthamiana) plants deficient in plant-specific α-1,3-fucosyltransferase and β-1,2-xylosyltransferase activity by mutating six different target genes (page 350, summery). It also teaches transformation of tobacco plants (page 358, left column, para 3) using expression vectors comprising polycistronic guide RNAs for editing genes involved in N-glycosylation (page 358, left column, para 3-4) and the gRNAs are transiently co-expressed with Cas9 in the host plant cells (page 351, right column, para 3, line 6-7). The Cas9 expression cassette comprising the Cas9 gene is cloned upstream of the gRNA cassette (bridging paragraph between page 351 and 352; page 352, Fig. 1).
Jansing et al. teaches a method by aligning exon sequences in different target genes to identify regions of homology that could be targeted by the same guide RNAs (gRNA) to knockout the target genes (page 351, right column, para 2). Using that method, Jansing et al. produces vectors containing 3-7 synthetic polycistronic gRNAs to achieve multiplex knockout of six genes (page 351, right column, para 2) encoding different isoforms of the enzymes viz. β-1,2-xylosyltransferase (XylT) and α-1,3-fucosyltransferase (FucT), involved in production of β-1,2-xylose and α-1,3-fucose in the plant. Jansing et al. also describes transforming the plants lacking N-glycosylation with a vector containing a gene encoding a monoclonal antibody 2G12 (summery; page 355, left column, para 2, line 1-3), which reads on to “a target protein”, as recited in claim 10.
However, Jansing et al. does not teach mutating or knocking out the genes in rice; or a polycistronic guide RNA expressing the nucleotide sequences of SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, and 15; or SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, and 8.
Steinkellner et al. (US 2020/0080100 A1) describes mutating in plant N-glycosylation genes encoding β1,2 – Xylosyltransferase; β1,3-Galactosyltransferases; α1,3-Fucosyltransferase; α1,4-Fucosyltransferase; and β Hexosaminidases (HEXOs) (page 6, para 0086). It also teaches use of CRISPR/Cas9 based gene editing system (page 6, para 0087, line 18-21) to introduce mutations in those genes to reduce or even abolish plant specific N-glycosylation (and resulting N-glycans) (page 6, para 0087, line 1-4). It also teaches use of rice plants (Oryza sativa) (page 23, para 0107) and rice cells for the invention (page 8, para 0107, line 5). Steinkellner et al. also describe mutations carried out using well-known and standard method of targeted genome editing technology using Cas9 (page 6, para 0087, line 18-20).
Kikuchi et al. teaches annotated cDNA sequences having 100% sequence identity to instant SEQ ID NOs: 1-9, 11, 13, and 15. Kikuchi et al. describes a cDNA clone (J013071F22, GenBank Accession No. AK099681) having 100% identity to SEQ ID NO: 1-2. The table below indicates cDNA clone numbers and GenBank Accession numbers, as taught by Kikuchi et al., having 100% sequence identity with specific SEQ ID NOs of this instant application.
SEQ ID NO.
Clone No. (Kikuchi et al.)
GenBank Accession No.
1
J013071F22
AK099681
2
J013071F22
AK099681
3
J033075J17
AK101954
4
J033075J17
AK101954
5
J013089J22
AK067033
6
J013089J22
AK067033
7
J023056D08
AK070606
8
J023056D08
AK070606
9
J023002J15
AK069009
11
J033108O18
AK121985
13
J023055K01
AK070632
15
J033093O08
AK102449
Before the effective filing date of this application, it would be obvious to one with ordinary skill in the art to make a plant expression vector to transform to mutate or knockout different genes involved N-glycosylation in a commercially important rice plant (as recited in claim 4) using CRISPR/Cas9 based gene editing system comprising a polycistronic guide RNA and a gene encoding Cas9, as taught by Jansing et al., by using the cDNA sequences, as taught by Kikuchi et al., to target endogenous genes encoding β1,2-Xylosyltransferase; β1,3-Galactosyltransferases; α1,3-Fucosyltransferase; α1,4-Fucosyltransferase; and β-Hexosaminidases (HEXOs), as taught by Steinkellner et al., with a realistic expectation to develop an easily scalable platform for the production of pharmaceutical proteins in rice wherein the plant-produced proteins would lack any plant specific N-glycosylation and no plant specific glycan. Genomic DNA sequence within the 1st or subsequent exon(s) of the target genes involved in N-glycosylation, viz. β1,2- Xylosyltransferase; β1,3-Galactosyltransferases; α1,3-Fucosyltransferase; α1,4-Fucosyltransferase; and β-Hexosaminidases (HEXOs) in rice, as taught by Kikuchi et al. would have enabled an ordinarily skilled artisan to design suitable polycistronic gRNAs to mutate or knocking-out the target genes to develop an easily scalable platform for the production of N-glycosylation deficient rice plants or rice cell lines.
Before the effective filing date, an ordinarily skilled artesian would have been motivated to make a plant expression vector to mutate or knockout various genes involved in N-glycosylation including genes encoding β1,2– Xylosyltransferase; β1,3-Galactosyltransferases; α1,3-Fucosyltransferase; α1,4-Fucosyltransferase; and β-Hexosaminidases (HEXOs) proteins in rice using a polycistronic guide RNA (gRNA) as part of CRISPR/Cas9 gene editing technique with a realistic expectation to develop an easily scalable platform for the production of pharmaceutical proteins in plants wherein the plant-produced proteins would lack any plant specific N-glycosylation and no plant specific glycan.
Response to Applicant’s Arguments
The argument set forth in the Applicant’s replies on 04/07/2026 has been fully considered but is not found persuasive.
Regarding claim rejections under 35 USC 103, the Applicant argues that “even if the expression of β1,2-XylT and α1,3-FucT is downregulated in rice in view of Jansing et al. and Steinkellner et al., β1,2-xylose and α1,3-fucose residues may be significantly reduced or not detected. Nevertheless, plant-specific glycan structures are not completely eliminated (page 7, para 3, line 1-4). The Applicant continued to argue, “knockout of ß1,2-XylT and α1,3-FucT does not block downstream B1,3-galactosylation and α1,4-fucosylation processes” (page 9, para 2, line 3-4) and still “is insufficient to achieve humanization of N-glycans, and additional modification of the N-glycosylation pathway to remove plant-specific residues such as ß1,3-galactose and α1,4-fucose is required (page 9, para 4, line 2-4). The Applicant mentions that “Steinkellner et al. does not provide any working examples in which all five genes are mutated simultaneously” (page 6, last para, line 1-2) and “Jansing et al. and Steinkellner et al. merely disclose mutating two genes, namely α1,3-FucT and β1,2-XylT, in tobacco. These references do not provide any working examples in which the β1,3-GalT and α1,4-FucT genes involved in N-glycosylation are mutated together with α1,3-FucT and β1,2-XylT, as in the present invention” (page 10, para 3, line 1-4).
The Examiner disagrees. Plant specific N-glycosylation is widely studied in the art. The enzymes and the genes encoding those enzymes are also known in the art, as taught by Kikuchi et al., before the effective filing date of the invention. Making vectors for various purposes including CRISPR-Cas based genome editing technique comprising suitable gRNA to make knockout mutants removing one or more specific genes is also a standard practice in the art (as described by Jansing et al. and Steinkellner et al., as discussed above), which can be used to mutate or remove one of more genes involved in N-glycosylation in plants including in both rice and tobacco, and is a routine standard process in the art. Moreover, not all (therapeutic) proteins need all different types of N-glycosylation present in a plant to produce a biologically active therapeutic protein when produced in a plant including in either rice or tobacco. There will be many therapeutic proteins (e.g. antibody 2G12), which would be biologically active in absence of just β-1,2-xylose and α-1,3-fucose in the plant producing the protein, as described by Jansing et al.
Moreover, Steinkellner et al. (US 2020/0080100 A1) describes mutations in all the genes involved in plant specific N-glycosylation, viz. the ones encoding β1,2 – Xylosyltransferase; β1,3-Galactosyltransferases; α1,3-Fucosyltransferase; α1,4-Fucosyltransferase; and all the β Hexosaminidases (HEXOs) in the plant that silence/inhibit/reduce their enzymatic activity (page 6, para 0086). It describes abolishing all plant specific N-glycans (page 6, para 0087, line 24-27). It teaches that the invention is equally applicable to rice plants (Oryza sativa) (page 23, para 0107) and rice cells (page 8, para 0107, line 5). Actual reduction to practice is not needed. Moreover, none of the claims recite anything to the effect of “completely removes plant-specific glycans” or “humanize N-glycan structure in rice”. Considering the growing importance to use various plant-based production platforms to produce growing number of therapeutic proteins with diverse glycosylation pattern, an ordinarily skilled artisan would have been motivated to mutate/knockout one or more specific gene(s) encoding one or more specific corresponding enzyme(s) involved in plant specific N-glycosylation to produce specific type of therapeutic proteins possessing required biological activity.
Applicant is reminded that the test for obviousness is not whether the features of a secondary reference may be bodily incorporated into the structure of the primary reference; nor is it that the claimed invention must be expressly suggested in any one or all of the references. Rather, the test is what the combined teachings of the references would have suggested to those of ordinary skill in the art. In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981).
Conclusion
No claim is allowed.
Communication
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/Jay Chatterjee/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
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