DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claims 1-7 are pending and under consideration.
Priority
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55 for Application No. CN2021116468045.5, filed on 12/29/2021.
Specification
The use of the term “OPTI-MEM” in paragraph [0057], which is a trade name or a mark used in commerce, has been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Claim Rejections - 35 USC § 112(b)
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.
Claim(s) 1-7 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.
Claim 1 is vague and indefinite in that the metes and bounds of the phrase “H4000 plasmid nanocarrier (Engreen),” are unclear. It is unclear whether the parenthetical phrase is a limitation or an example of a company from which the nanocarrier can be purchased. It would be remedial to delete “(Engreen)” from the claim.
Claim Rejections - 35 USC § 112 (Written Description)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim(s) 1 and 3-7 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The fundamental factual inquiry is whether the specification conveys with reasonable clarity to those skilled in the art that, as of the filing date sought, Applicant was in possession of the invention as now claimed. See, e.g., Vas-Cath, Inc., 935 F.2d at 1563-64, 19 USPQ2d at 1117.
Claim(s) 1 and 3-7 are drawn to a genus of dCas9 plasmid sequence for enhancing the expression of demethylase TET2. The rejected claims thus comprise a genus of dCas9 plasmid sequences and are defined as belonging to the broad class of dCas9 and as having the function of enhancing the expression of demethylase TET2.
To satisfy the written description requirement, MPEP §2163 states, in part “… a patent
specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention.” Moreover, the written description requirement for a genus may be satisfied through sufficient description of a representative number of species by “… disclosure of relevant, identifying characteristics, i.e.,
structure or other physical and/or chemical properties, by functional characteristics coupled with
a known or disclosed correlation between functional and structure, or by a combination of such
identifying characteristics, sufficient to show the applicant was in possession of the claimed genus.”
The specification envisions the dCas9 plasmid sequence as, “In some embodiments, the dcas9 plasmid sequence is pZdonorU6-sgRNA-EFla- dSpCas9-NLS -VP64-2A-EGFP-2A-Puro.”, (see paragraph [0011]; Fig. 1A; SEQ ID NO: 1). Moreover, the specification describes the use of the plasmid as, “After the Crispr/dcas9 system is transfected into Treg cells, the expression of a TET2 gene promoter is enhanced, and the expression of TET2 protein is upregulated, thereby regulating the secretion of Treg cell-related cytokines, and further promoting the nerve reconstruction of Crispr/dcas9-modified engineered blood vessels.”, (see paragraph [0023]).
Even if one accepts that the examples described in the specification meet the claim limitations of the rejected claims in regard to structure and function, the examples are only representative of one dCas9 plasmid used for enhancing the expression of TET2 (SEQ ID NO:1 containing VP64). These results are not necessarily predictive of all “dCas9 plasmid sequences” capable of enhancing expression of TET2. Thus, it is impossible for one to extrapolate from the one example of dCas9 plasmid sequence described herein that the “dCas9 plasmid sequence” would necessarily meet the structural/functional characteristics of the rejected claims.
The prior art does not appear to offset the deficiencies of the instant specification in that it does not describe a set of “dCas9 plasmid sequence” capable of enhancing the expression of TET2.
Looking to a review article on CRISPR/Cas9 therapeutics, Li et al (CRISPR/Cas9 therapeutics progress and prospects, Signal Transduction and Targeted Therapy, Vol 8, Issue 36, Pages 1-23, published January 16th, 2023) describes Cas9 and dCas9 structure:
Li et al discloses in ‘The transcriptional regulatory tool dCas9’ section, “The DNA strand cleavage function of Cas9 was elucidated by designing a simple sgRNA segment to guide Cas9 to the target site, but many additional studies on genes have been performed to address functions other than DNA strand cleavage. Qi et al. mutated the RuvC1 and HNH nuclease domains (D10A and H841A) of the wild-type Cas9 mentioned above, causing Cas9 to lose its cleavage enzyme activity. dCas9 showed efficient gene silencing when sgRNAs were designed for nontemplate DNA strands, while sgRNAs designed for template strands did not effectively silence gene expression. The relative positions of sgRNAs and target gene promoter sequences also had a significant effect on silencing efficiency. Importantly, for the sgRNA targeting promoter sequences, gene silencing occurs regardless of whether the target is the template or nontemplate strand. In July 2013, another study by Qi et al. revealed that dCas9 interacts with effectors related to transcriptional regulation, such as VP64 and KRAB, to coregulate gene expression, which is currently the most common use of dCas9.”, (see page 3, column 2, paragraph 3).
More specifically Li et al discloses in the ‘Validity’ section, “The initial transcriptional activation domain is the VP64 or p65 activation structural domain formed by the complex of four transcribed VP16, and the activation of this structure is not strong. Tanenbaum et al. constructed a synthetic system composed of a structure that contains a polypeptide chain that can recruit up to 24 copies of the protein to obtain higher activation efficiency. The structure was used to recruit multiple copies of VP64 to form the dCas9-SunTag-VP64 transcriptional activation system. In a study of the activation of cell cycle suppressor cyclin-dependent kinase inhibitor 1B (CDKN1B), dCas9/VP64 did not affect cell cycle progression, whereas the same sgRNA carrying dCas9-SunTag-VP64 significantly inhibited cell cycle progression and reduced cell growth. VP64, p65 and Rta have been reported to have the ability to activate transcriptional. Chavez et al. used dCas9/VP64 as a backbone and added p65 and Rta to construct the transcriptional complex dCas9-VP64-p65-Rta (dCas9/VPR) as a transcriptional activation system. With its simple structure and high efficiency, dCas9/VPR is one of the most frequently used activation systems for targeted delivery of CRISPRa. Other transcriptional activation systems, such as dCas9/SAM, dCas9/SPH and dCas9/VP192, are also able to substantially increase the efficiency of gene activation.”, (see page 16, column 2, paragraph 1; and Table 4).
Therefore, the art does not appear to offset the deficiencies of the specification. Merely describing a “dCas9 plasmid sequence” capable of enhancing expression of TET2 without sufficient detail relating to the genus of dCas9 plasmid sequence in enhancing expression of TET2 does not allow the skilled artesian to reasonably conclude that the Applicants were in possession of the claimed invention in claim(s) 1 and 3-7.
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.
Claim(s) 1-7 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Wang et al (Programming of Regulatory T Cells In Situ for Nerve Regeneration and Long-Term Patency of Vascular Grafts. Research. Vol 2022, Issue 9826426, Pages 1-17, July 25th, 2022).
Applicant cannot rely upon the certified copy of the foreign priority application to overcome this rejection because a translation of said application has not been made of record in accordance with 37 CFR 1.55. When an English language translation of a non-English language foreign application is required, the translation must be that of the certified copy (of the foreign application as filed) submitted together with a statement that the translation of the certified copy is accurate. See MPEP §§ 215 and 216.
Regarding Claim 1, Wang et al discloses implantation of CRISPR/dCas9 nanodelivery system-modified sdTEVGs in rats that resulted in Treg cell editing, control of excessive inflammation, and promoted nerve regeneration (abstract). More specifically, Wang et al discloses, “To enhance TET2 expression in Treg cells in vivo and in vitro, a nonviral plasmid DNA delivery system containing an enhanced TET2 gene promoter named as dcas9 plasmid, cationic polymers H4000 as transfection vector, and targeting peptides was designed (Figures 2(a)–2(c))… Flow cytometry revealed that the transfection (Foxp3 and GFP positive Treg cells) and activation efficiencies (CD44 + GFP + Treg cells) were significantly increased after modification of the CRISPR/dCas9 nanodelivery system with the CD25 antibody (Figure 2(d)-2(g))… These results indicated the superior targeting of the CD25 antibody-modified NPs (H4000-CD25/dCas9) to that of the unmodified NPs (H4000/dCas9). The CD25 antibody enhanced the targeted editing efficiency of the gene delivery system.”, (Page 2, Column 2, Paragraph 2).
Moreover, Wang et al discloses, “2.4. Design and Properties of sdTEVGs Modified with H4000-CD25/dCas9 Nanodelivery System. Based on these results, the protonated H4000-CD25/dCas9 delivery system and collagen with many electrons were used to construct a long term sustained-release engineering vessel using an electrostatic layer-by-layer self-assembly method (Figures 4(a) and 4(b))…. After encapsulating the plasmid, the H4000-CD25/dCas9 system appeared in clusters, and the sdTEVGs modified with H4000-CD25/dCas9 showed that many nanoparticles were adsorbed onto the adventitia surface (Figures 4(c) and 4(f)).”, (Page 5, Column 2, Paragraph 1).
Regarding claim 2, Wang et al discloses the dCas9 plasmid sequence under section 4.5 dCas9 Plasmid Design: The transfection vector was constructed and synthesized by Beijing Syngen-Tech Co. as follows: pZdonor_U6-sgRNA-EF1α-dSpCas9-NLS-VP64-2A-EGFP-2A-Puro (Page 12, Column 1, paragraph 3; and Figure 2a).
Regarding claim 3, Wang et al discloses under section 4.20.1. SdTEVG Transplantation: All sdTEVGs (length:1 cm, diameter: 1mm) in the H4000-CD25PE, H4000/dCas9, and H4000 CD25PE/dCas9 groups were transplanted into the carotid artery of 6–8-week allogenic SD rats, which were injected intraperitoneally with heparin sodium (1,000Ukg-1, Aladdin) for 4 days, and maintained in the absence of a specific pathogen (Page 14, Column 1, Paragraph 2). Wherein sdTEVGs are small diameter tissue engineering vascular grafts (abstract). Figure 4a also discloses sdTEVG as tubular.
Regarding claim(s) 4-7, Wang et al teaches a method for preparing the cardiovascular implant based on in-situ regulation of immune response.
Claim 4 sets forth steps for preparing the cardiovascular implant: Step 1, constructing a cardiovascular implant body and step 4, conjugating the H4000-CD25/dcas9 sustained-release nanoparticles on the cardiovascular implant body: co-incubating the cardiovascular implant body with the H4000-CD25/dcas9 sustained-release nanoparticles and collagen to obtain the cardiovascular implant, (which is further limited by claim 5 and claim 7). Wang et al discloses the claimed steps of claim 5 and 7 in Figure 4A and methods section, “4.16 Preparation of sdTEVGs. Under sterile conditions, the carotid arteries of SD rats were harvested and decellularized with 0.05% trypsin (Gibco) at 37°C for 30 min and washed thrice with PBS for 5 min. Subsequently, acellular blood vessels were soaked in h4000-cd25pe, h4000/dCas9, and h4000-cd25pe/dCas9 for 10 min, washed with PBS for 2 min, and again soaked in a soluble collagen PBS solution (1 g L-1) for 10 min. SdTEVGs were obtained after repeating the incubation and elution thrice.”, (Page 13, Column 2, Paragraph 2).
Step 2, preparing an H4000-CD25/dcas9 nanotransfection vector is disclosed by Wang et al in methods section “4.6. Manufacturing of H4000-CD25PE and H4000-Cy3 Nanoparticles. The Entranster-H4000 (H4000, Engreen) was diluted from 20 μL to 1 mL. Hydrodynamic dimension and zeta potential were measured. CD25-PE (100 μL,0.2mgmL-1) was mixed with EDC (300 μL, 1mgmL-1) in MES (pH 5.5), and then, H4000 (100 μL) was added overnight
in the dark. Finally, H4000-CD25PE was purified using a dialysis bag with a molecular weight of 100kD and enriched in 100 μL of water. The hydrodynamic force and zeta potential of the product were measured again to check for coupling effect, and 20 μL was used to perform the fluorescence spectrometer test. Activated Cy3-NHS (10 μL, 5mgmL-1), H4000 (200 μL), and NaHCO3 (200 μL, 0.1 M, pH7.8) were homogeneously mixed and reacted overnight in the dark at 4°C. Subsequently, the reacted solution was dialyzed (molecular weight: 3500 D) in deionized water for 24 h, and the H4000-cy3 nanoparticles were obtained. Furthermore, H4000 and purified H4000-CY3 were diluted to 0.5mgmL-1 with deionized water, and the hydration particle
size and surface potential of each sample were tested thrice using a laser particle size analyzer (Malvern, zen3690).”, (Page 12, Column 1, Paragraph 4).
Lastly, step 3, preparing H4000-CD25/dcas9 sustained-release nanoparticles by the H4000-CD25 nanotransfection vector and a Crispr/dcas9 system plasmid (which is further limited by claim 6) is disclosed in Wang et al. Wang et al discloses in methods section, “4.7. dCas9 Plasmid Packaging in Entranster-H4000. The dCas9 plasmid and vector Entranster-H4000/H4000-CD25PE were diluted with serum-free RPMI to a working concentration of 32 μg mL-1 and 80 μg mL-1. To prepare the transfection complex, the Entranster-H4000 was mixed with the plasmid at a volume ratio of 2:5 and kept for 15 min at 18–25°c. The following groups
of transfected vectors and plasmids were mixed using the same methods: H4000-CD25PE, H4000/dCas9, and H4000-CD25PE/dCas9.”, (Page 12, Column 1, paragraph 5 to Column 2, Paragraph 1). Wherein, “To prepare the transfection complex, the Entranster-H4000 was mixed with the plasmid at a volume ratio of 2:5…” of Wang et al reads on the “wherein the Crispr/dcas9 system plasmid is incubated with the H4000-CD25 nanotransfection vector at 0.4 pg of plasmid/pL of transfection vector.”, of claim 6.
Thus, claims 1-7 are anticipated by Wang et al.
Conclusion
No claims are allowed.
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/L.M.T./Examiner, Art Unit 1637
/Jennifer Dunston/Supervisory Patent Examiner, Art Unit 1637