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 .
Election/Restrictions
Applicant's election with traverse of the Species election between CRISPR/Cas13a and microRNA expression vector in the reply filed on 02 October 2025 is acknowledged. The traversal is on the ground(s) that the claimed invention of synergistic cooperation of DMP-controlled inhibition of iron export genes (FPN/LCN2) and iron nanoparticles is not obvious. This is not found persuasive because of the rejection set forth below.
The requirement is still deemed proper and is therefore made FINAL.
Application Status
The Amendments and Remarks filed 10 October 2025 in response to the Office Action 03 July 2025 are acknowledged and have been entered. Claims 1, 3-10, and 13 are amended,. Claims 2 and 11 have been cancelled. Claims 1, 3-10, and 13 are pending and being examined on the merits.
Priority
This application is a 371 PCT of CN2021/072025 filed 01/15/2021 and claims priority to application CN202010173472.2 filed 3/12/2020.
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55. Should applicant desire to obtain the benefit of foreign priority under 35 U.S.C. 119(a)-(d) prior to declaration of an interference, a certified English translation of the foreign application must be submitted in reply to this action. 37 CFR 41.154(b) and 41.202(e).
Failure to provide a certified translation may result in no benefit being accorded for the non-English application.
Information Disclosure Statement
The information disclosure statement filed 09/12/2022 has been acknowledged.
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
This application contains sequence disclosures in accordance with the definitions for nucleotide and/or amino acid sequences set forth in 37 CFR 1.831(a) and 1.831(b). However, this application fails to comply with the requirements of 37 CFR 1.831-1.834. The examiner has noted that claim 9 and the specification recites nucleic acid sequences that lack corresponding SEQ ID NOs. Applicant must provide:
• A replacement “Sequence Listing XML” part of the disclosure, as described above in item 1. or 2., as well as
• A statement that identifies the location of all additions, deletions, or replacements of sequence information in the “Sequence Listing XML” as required by 1.835(b)(3);
• A statement that indicates support for the amendment in the application, as filed, as required by 37 CFR 1.835(b)(4);
• A statement that the “Sequence Listing XML” includes no new matter in accordance with 1.835(b)(5); and
• A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3), and 1.125 inserting the required incorporation by reference paragraph as required by 37 CFR 1.835(b)(2), consisting of:
o A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
o A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Specification
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code in paragraphs 0037, 0102, and 0104. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
The use of the term Sangon Biotech, Thermo Fisher Scientific, Cell Counting Kit-8, BioTek, Lipofectamine 2000, CytoFLEX LX, Beyotime, Beckman, Signalway Antibody, Licor, Abcam, Odyssey, Changzhou Cavens Laboratory Animal Co, Invitrogen, TIANGEN, Yeasen, Applied Biosystems, GraphPad Prism, GraphPad Prism, to name a few, 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. A cursory review of the specification has revealed these trade names or marks. It would be remedial to identify and amend all trade names or marks in the specification upon amendment
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 Objections
Claim 3 is objected to because of the following informalities:
Claim 3 recites “ wherein the cancer cell specific promoter DMP is a NF-κB-specific promoter”. While it is clear that this recitation is referring to the decoy minimal promoter (DMP) of claim 1, it would be remedial recite this phase to say “ wherein the DMP promoter is a cancer cell NF-κB-specific promoter.
Claim 3 seems to be incomplete by ending in the term “in”.
Claim 3 recites “κB decoy”. It would be remedial to change this to “NF-κB decoy”.
Appropriate correction is required.
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 1, 3-10, and 12 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 recite, “the cancer cell” in line 10. There is insufficient antecedent basis for this limitation in the claim.
Claim 1 provides for a gene interference vector (CRISPR/Cas13a or microRNA expression vector) that expresses a (i.e., one) gRNA/microRNA that inhibits expression of an iron export gene selected from FPN and LCN2. Claim 1 further recites that “the Cas13a-gRNA or microRNA… inhibits expression of one or more intracellular iron metabolism genes or reactive oxygen species-related genes”. Claim 1 only provides for one gene interference expression vector and one gRNA/microRNA; so it is unclear how the vector inhibits expression of both an iron export gene and one or more intracellular iron metabolism genes or reactive oxygen species-related genes. Furthermore Fig. 3 of the specification teaches that the Cas13a-gRNA or mircoRNA inhibits FPN and LCN2. For the purpose of examination, the claims will be examined based on the gene interference vector inhibiting only an iron export gene selected from FPN and LCN2.
Claim 3 recites that the NF-κB-specific promoter is composed of NF-κB. A promoter is a composed of nucleotides and NF-κB is a protein composed of amino acids. It is unclear how a nucleotides can be composed of amino acids.
Those claims identified in the statement of rejection but not explicitly referenced in the rejection are also rejected for depending from a rejected claim but failing to remedy the indefiniteness therein
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 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 nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1, 3-4, 6, 8, 10, and 12 are rejected under 35 U.S.C. 103 as being unpatentable over Chen (Chen et al. ACS Biomater. Sci. Eng. 2019, 5, 4861−4869), Kong (Kong et al. Cell Death and Disease (2019) 10:624), Pellenz (US 2021/0261960 A1, foreign filed 2/26/2020, and Southeast University (CN108220336A, published 06/29/2018).
Regarding claim 1, 3-4, 8, 10, Chen teaches the design method of using of ultrasmall poly(ethylene glycol)-modified polydopamine nanoparticles, (UPDA-PEG)@ Fe2+/3+ nanoparticles (iron NPs) for ferroptosis-induced tumor cell death [abstract; Fig. 1, Scheme 1]. Chen teaches that these nanoparticles release of iron ions can reach approximately 70% at pH 5.0, providing the advantage of application in tumor microenvironment [abstract, Fig. 2C]. Chen teaches that iron nanoparticles increase ROS [pg. 4865, col.1, para 3; Fig. 2d]. Chen teaches that the administration of iron NPs in tumor-bearing Balb/c mice could significantly inhibit tumor progression [abstract]. Chen teaches that the iron NPs reported herein represent the nanoparticles related to iron ions for chemotherapy against cancer through the ferroptosis pathway [abstract].
Chen does not teach wherein the method additionally comprises administering a CRISPR/Cas13a gene interference expression vector under the control a decoy minimal promoter (DMP), wherein said vector expresses a gRNA that inhibits expression of an iron export gene selected from FPN wherein the inhibition of the iron export gene prevents efflux of intracellular iron ions.
Kong teaches that Nrf2-mediated FPN1 downregulation promoted intracellular iron accumulation and reactive oxygen species [abstract]. Kone teaches the use of Cas9/sgRNA targeting FPN1 for the knockout of FNP1 [pg. 8, col. 1, para 4]. Kong teaches that knocking out FPN1 resulted in higher intracellular iron levels, compared with Cas9/sg Ctrl treatment and higher ROS levels [pg. 8, col. 1, para 5].
Pellenz teaches unlike Cas9, Type VI CRISPR associated endonucleases like Cas13a recognize ssRNA rather than dsDNA [0010-0011]. Pellenz teaches that by using RNA CRISPR-Cas13a ribonucleoproteins in mammalian cells, knockdown levels have been attained comparable to RNAi, but with improved specificity [0011]. Pellenz teaches that components of the CRISPR Cas Systems can be delivered using vectors or AAV particles [0249, 0231, 0235].
Southeast University develops a NF-κB specific activation gene expression vector, which could induce the death of cancer cells in vitro and in vivo by utilizing NF-κB over-activity in cancer cells, but had no effects on normal cells [abstract; pg. 5, para 1 (Fig. 9); pg. 7, para 10]. Southeast University teaches that the vector contains a NF-κB response promoter (DMP) composed of NF-κB response sequence and a minimal promoter, which can control the specific expression of downstream effector genes in NF-κB overactivated cells [pg. 4, para 4]. Southeast University teaches the vector could be used to express CRISPR/Cas9 protein and sgRNA only in cancer cells, where the Cas9 protein was then guided by a sgRNA targeting telomeric DNA and induced cancer cell death but not normal cells [pg. 8, para 1]. Southeast University teaches the cell introduction method is a viral vector; more preferably, adeno-associated virus (AAV) [pg.3, para 14; pg. 4, para 4]. Southwest University teaches the U6-TsgRNA-DMP-Cas9 expression vector of which the sgRNA is controlled by the U6 promoter [pg. 5; para 3; Fig. 11].
Regrading claim 12, Chen teaches delivering the iron NPs as a chemical material and Southeast University and Pellenz teaches that the Cas/sgRNA vector can be delivered as an AAV vector.
It would have been obvious to one ordinary skilled in the art before the effective filing date of the claimed invention to modify the method of Chen by additionally including a CRISPR/Cas13a gene interference expression vector under the control a decoy minimal promoter (DMP), wherein said vector expresses a gRNA that inhibits expression of an iron export gene selected from FPN wherein the inhibition of the iron export gene prevents efflux of intracellular iron ions. One of ordinary skill would be motivated to make the modification for the advantage of killing cancer cells. Furthermore, the combination of prior art elements according to known methods to yield predictable results supports can support a conclusion of obviousness. See MPEP 2143(I). One of ordinary skill in the art would have a reasonable expectation of success since Chen teaches that an intracellular increase in iron ions and ROS can be used as chemotherapy against cancer through the ferroptosis pathway; Kong teaches that the CRISPR/Cas mediated downregulation of FPN1 leads to an increase of iron ions and ROS, and Southeast University teaches that a DMP, NF-KB,-specific promoter can drive the expression of Cas proteins for the advantage of killing cancer cells.
Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over Chen (Chen et al. ACS Biomater. Sci. Eng. 2019, 5, 4861−4869), Kong (Kong et al. Cell Death and Disease (2019) 10:624), Pellenz (US 2021/0261960 A1, foreign filed 2/26/2020, and Southeast University (CN108220336A, published 06/29/2018) as applied to claim 1, and further in view of GenBank: AF226614.1 (GenBank: AF226614.1. Homo sapiens ferroportin1 (FPN1) mRNA, complete cds. 2/29/2000).
The teachings of Chen, Kong, Pellenz, and Southeast University are discussed above as applied to claim 1, and similarly apply to claim 9.
Chen, Kong, Pellenz, and Southeast University do not teach where the target binding sequences of the gRNAs targeting FPN is 5'-CACCG CAAAG TGCCA CATCC GATCT CCC- 3'.
GenBank: AF226614.1 teaches the complete cds for the Homo sapiens ferroportin1 (FPN1) mRNA. GenBank: AF226614.1 teaches residues 415-442 which is 100% identical to 5'-CACCG CAAAG TGCCA CATCC GATCT CCC- 3'.
It would have been obvious to one ordinary skilled in the art before the effective filing date of the claimed invention to use the sequence of GenBank: AF226614.1 as gRNA targeting sequence of FPN. One of ordinary skill would be motivated to used this as the gRNA targeting sequence since this sequence was known as an mRNA sequence of FPN.
Allowable Subject Matter
The following is a statement of reasons for the indication of allowable subject matter: SEQ ID NO: 1 is found free of the art. SEQ ID NO: 1 is taught by the specification to the CRISPR/Cas13a expression vector (pDMP-Cas13a-U6-gRNA). While SEQ ID NO: 1 comprises the Southeast University’s NF-κB response sequence (5'-GGG AAT TTC CGG GGA CTT TCC GGG AAT TTC CGG GGA CTT TCC GGG AAT TTCC-3', SEQ ID NO.1) and the minimal promoter sequence (5'-TAG AGG GTA TAT AAT GGA AGC TCG ACTTCC AG- 3', SEQ ID NO.2) of the DMP promoter, comprises a sequence that is 99.6 identical to GenBank: AY623053.1 (GenBank: AY623053.1 Homo sapiens clone phU6A U6 small nuclear RNA, promoter region, 9/7/2005) who teaches a U6 promoter region, and comprises a sequence that is 90% identical to GenBank: MN812663.1 (GenBank: MN812663.1, Vector pAc-Cas13a, complete sequence, 2/29/2020), the prior art failed to teach and suggest arranging the components of a DMP promoter, Cas13a, U6 promoter, and a gRNA to arrive at SEQ ID NO:1.
Claim 5 is objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening claims.
Conclusion
No claims allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to TIFFANY N GROOMS whose telephone number is (571)272-3771. The examiner can normally be reached M-F 830-530.
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/TIFFANY NICOLE GROOMS/Examiner, Art Unit 1637