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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 11/18/2025 has been entered.
The objection to the Specification has been withdrawn in view of Applicant’s amendments.
The rejection under section 103 is withdrawn in favor of the following new ground of rejection:
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 20-22, 24-27, 30-34, 37, 39-42, 68-72 are rejected under 35 U.S.C. 103 as being unpatentable over:
U.S. Publication No. 20160090603 based on an application by Carnes et al. (Carnes); or
Lipofectamine 2000 Reagent, 2013, downloaded 29 November 2024 from https://assets.thermofisher.com/TFS-Assets/LSG/manuals/Lipofectamine_2000_Reag_protocol.pdf (Lipofectamine); or
Zuris et al., Nat Biotechnol. 2014 Oct 30;33(1):73–80, newly cited (Zuris);
in view of:
Mahato, Advanced Drug Delivery Reviews 57 (2005) 699 – 712 (Mahato).
Paragraph 180 of Carnes teaches ribonucleoprotein complexes comprising ‘lipoplexes’, which are defined by Carnes as comprising cationic lipids and genetic material, CRISPR components and enhancer peptides.
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Carnes specifically contemplates lipid packages including nucleases:
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The lipid layer comprises cationic lipids are discussed, see paragraphs 0146+.
Carnes teaches eukaryotic host cells:
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Zuris teaches that common cationic lipid nucleic acid transfection reagents can mediate the potent delivery of Cas9:sgRNA nuclease complexes into cultured human cells in media containing 10% serum, see Abstract.
The primary references may not explicitly teach an enhancer element that comprises a polyamine moiety covalently attached to a nuclear localization signal sequence, wherein said polyamine moiety comprises a spermine moiety. However, it is for that proposition that the examiner joins the secondary references which teach eukaryotic transfection and nuclear localization with the recited enhancer elements:
Mahato demonstrates that Lipofectamine is cationic lipid available for gene therapy/editing (2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propanammonium trifluoroacetate (DOSPA)), see page 700.
In this way, those of ordinary skill could have applied the recited enhancer elements in the manner required and in a predictable fashion for the purposes of obtaining the recited complexes. The primary references teach lipoplexes comprising protein and nucleic acids. The secondary references are added for the proposition that the recited enhancer element is applicable to these complexes. Specifically, Mahato teaches that the particular known technique of using the recited enhancer elements to facilitate transfection and nuclear localization of ribonucleoproteins was recognized as part of the ordinary capabilities of one skilled in the art. In this manner, those of ordinary skill would have recognized that applying the known technique to those methods that utilize transfection of ribonucleoproteins, such as the CRISPR reagents taught by the primary references, would have yielded predictable results. Accordingly, using the recited enhancer elements with the lipoplexes of the primary references to provide a nucleic acid-protein transfection agent would have been prima facie obvious.
Applicant argues that the references fail to teach lipid-based eukaryotic cell transfection in an aqueous solution.
However, Carnes exemplifies particles in Figs. 1A-1D:
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Carnes specifically teaches that any silica surface can be optionally modified with biocompatible lipids to increase the colloidal stability of NanoCRISPRs and to facilitate their conjugation with ligands that target particular cells or organelles within the host cell or that promote endosomal escape of NanoCRISPRs upon host cell uptake. (D) Also shown is a schematic of a NanoCRISPR delivery platform (e.g., a protocell or a silica carrier) interacting with the host cell to deliver the biological package. (1) Targeting ligands conjugated to the NanoCRISPR surface can bind to corresponding receptors on the host cell. (2) Binding can trigger receptor-mediated endocytosis of NanoCRISPRs. (3) Endosomes become acidified, which will cause the lipid coating to dissociate from the NanoCRISPR's silica surface, see paragraph 0060.
As outlined above, Zuris teaches that common cationic lipid nucleic acid transfection reagents can mediate the potent delivery of Cas9:sgRNA nuclease complexes into cultured human cells in media containing 10% serum.
Accordingly, lipid-based eukaryotic cell transfection in an aqueous solution was known and is prima facie obvious.
The following is a new ground of rejection, which was necessitated by Applicant’s amendments:
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 20-22, 24-27, 30-34, 37, 39-42, 68-72 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
The structural criteria of “lipid-based eukaryotic cell transfection composition” is not defined.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to KARL J PUTTLITZ whose telephone number is (571)272-0645. The examiner can normally be reached on Monday to Friday from 9 a.m. to 5 p.m.
If attempts to reach the examiner by telephone are unsuccessful, the examiner's acting supervisor, Janet Epps-Smith, can be reached at telephone number (571)272-0757. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/KARL J PUTTLITZ/ Primary Examiner, Art Unit 1646