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 of Group I (claims 1, 2, 4, 7, 10, 16, 18, 29, 45, and 53) is acknowledged and accepted for examination. Applicant’s election of species, including (i) trnl as a species of first flanking sequence and trnA as a species of second flanking sequence (from claim 2); (ii) item (b) from claim 4, and from between rpoC2 and rps2 genes as a species of spacer; (iii) item (a) from claim 7 and psbA gene as a species of promoter; (iv) item (c) from claim 10, and T7GI0 as a species of 5' UTR and a heterologous psbC gene as a species of 3' UTR;
(v) item (c) from claim 16, and Staphylococcus aureus Protein A as a species of a protein of interest; (vi) a plastid from a dicot plant, where the dicot plant is a low-nicotine tobacco plant (from claim 18); (vii) item (b) from claim 29; and (viii) item (c) of claim 45, is also acknowledged.
The traversal of the restriction requirement between Group I and Group II has been considered but is not persuasive. The claims of Group I and Group II are directed to distinct inventions, and Group I is drawn to a transformation vector for plastid transformation, whereas Group II is directed to an expression cassette, which constitutes a sperate and distinct invention. Accordingly, the restriction requirement between Groups I and II is maintained.
Examination on the merits is limited to the elected group.
Claim Status
Claims 1, 2, 4, 7, 10, 16, 18, 29, 45, and 53 are pending.
Claims 1, 2, 4, 7, 10, 16, 18, 29, 45, and 53 are examined on the merits.
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 4 and 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 4 is rejected as indefinite for the recitation “….the first and second flanking sequences are substantially homologous to sequences…”. The term of “substantially homologous” lacks an objective boundary. The claim does not specify the degree of sequence identity or permissible variation required, such that the metes and bounds of the claimed flanking sequences are not reasonably certain. Claim 4, for recitation that “the spacer is from between” the identified plastid genes, is indefinite because it is unclear whether the claimed spacer must be limited to the intergenic region between those genes or may include part of one or both adjacent coding regions. Thus, the boundaries of the claimed spacer are not reasonably certain.
Claim 7, for reciting that “the promoter is a mutated 16S rRNA promoter with reduced homology to the endogenous 16S rRNA promoter yet with substantially equal functionality”, is indefinite for two reasons. First, the term “substantially equal functionality” lacks an objective standard, as the claim does not provide a benchmark for determining what level of equivalence is required. Second, it is unclear what “functionality” refers to, i.e., whether it pertains to promoter activity, transcriptional strength, or another function. Accordingly, the scope of the claim is not reasonably certain.
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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1 and 4 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Maliga (Pal Maliga et. al., US5877402A, Application: 1994-01-31, Publication: 1999-03-02).
Claim 1 recites a transformation vector for stably transforming a plastid, said transformation vector comprising: an expression cassette comprising, as operably-linked components, a regulatory sequence operative in the plastid, and a heterologous polynucleotide sequence coding for a protein of interest, and flanking each side of the expression cassette, a first DNA flanking sequence and a second flanking DNA sequence which allow for stable integration of the heterologous polynucleotide sequence coding for the protein of interest into the plastid genome.
Maliga discloses a transformation vector for stably transforming a plastid (claim 1), comprising a transforming DNA construct/vector having a targeting segment with plastid DNA homologous to a predetermined plastid genomic sequence, thereby enabling insertion into the plastid genome by homologous recombination (claim 1a). Maliga further discloses a selectable marker gene and at least one additional expressible DNA segment disposed within the targeting segment, wherein the expressible DNA segment contains a coding segment encoding a gene product (claim 1bi). Maliga further discloses targeting insertion of heterologous sequence with homologous plastid DNA insertion (column 21, line 30-45). Maliga discloses a 5’regulatory segment of plant chloroplast origin operably linked to the coding segment to promote expression in plastid (claim 1bii), and a 3’regulatory segment of a plant chloroplast mRNA molecule operably linked to the coding segment to promote stability of mRNA produced during expression (claim 1biii).
Thus, Maliga discloses an expression cassette comprising, as operably linked components, a regulatory sequence operative in the plastid and a heterologous polynucleotide sequence coding for a protein of interest.
Maliga also discloses that the construct includes plastid DNA sequence flanking the expression elements which enable stable integration into the plastid genome by homologous recombination (claim 1a and claim 3a), thereby discloses flanking each side of the expression cassette, a first DNA flanking sequence and a second DNA flaking sequence which allow for stable integration of the heterologous polynucleotide sequence coding for the protein of interested into the plastid genome.
Accordingly, Maliga discloses every limitation of claim 1, and claim 1 is anticipated by Maliga.
Under BRI, Claim 4 recites the vector of claim 1, wherein: (a), (b), and (c) in the alternative. For purpose of this rejection, claim 4 is examined with respect to subpart (a) only, which recites the first and second flanking sequences are substantially homologous to sequences around an integration site of the plastid genome and provide for homologous recombination to insert the heterologous polynucleotide coding for the protein of interest into the integration site of the plastid genome.
As set forth for the same reasons as claim 1, Maliga discloses a transformation vector comprising an expression cassette and flanking sequences that allow for stable integration into the plastid genome. Further, Maliga discloses that the flanking sequences are homologous to plastid genomic sequences and mediate homologous recombination to insert the heterologous polynucleotide into the plastid genome (claim 1, column 21, line 30-45).
Therefore, Maliga discloses the limitations of claim 4(a), and claim 4 is anticipated.
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 2, 7, and 10 are rejected under 35 U.S.C. §103 as being unpatentable over Maliga (US5877402A) as applied claim1, in view of Lutz (Kerry Ann Lutz et. a., Plant Physiology (2007)145(4):1201-10).
Claim 1 as the teaching of Maliga is discussed above.
Claims 2, 7, and 10 are interpreted as depend of claim 1.
Claim 2 recites the vector of claim 1, wherein (a) the first flanking sequence comprises trnI as species election, and (b) the second flanking sequence comprise comprises trnA as species election.
Maliga teaches plastid transformation construct comprising plastid flaking sequence for homologous recombination into the plastid genome (claim 1 and 3). Maliga does not teach trnI as first flanking sequence and trnA as second franking sequence.
Lutz teaches plastid vector targeting insertion in the trnI-trnA region (page 1201, Abstract). It would have been obvious for a person having ordinary skill in the art to use the known trnI and trnA flanking sequences in the plastid construct of Maliga as a known plastid insertion site for targeted plastid integration.
Accordingly, claim 2 is obvious over Maliga and Lutz.
Claim 7 recites the vector of claim 1, wherein the regulatory sequence comprises a promoter operative in the plastid genome, and wherein: a) the promoter is selected from psbA gene as species election.
Lutz teaches use of plastid psbA promoter in plastid expression cassettes (page 1202, right collum paragraph 3, and page 1205 Table 1). It would have been obvious to use the known plastid psbA promoter in the plastid construct of Maliga for plastid expression of the heterologous coding sequence. Accordingly, claim 7 is obvious over Maliga and Lutz.
Claim 10 recites the vector of claim 1, wherein the regulatory sequence further comprises:(a) a 5' untranslated region (UTR) capable of providing transcription and translation enhancement of the heterologous polynucleotide coding for the protein of interest, wherein the 5' UTR is (i) a 5' UTR of T7G10 as species election; (b) a 3' UTR capable of conferring stability to a transcript of the protein of interest, wherein the 3' UTR is (i) a 3' UTR of psbA gene.
Maliga teaches plastid 5’regulateory segments and plastid 3’regulatory segments for promoting expression and mRNA stability in plastids (claim 1bii and 1biii), and Lutz teaches the T7 gene 10 leader for high-level translation in chloroplasts (page 1206, left column paragraph 1) and plastid 3’UTR regulatory elements including psbA (page 1202, right column, paragraph 3; page 1205, Table 1).
It would have been obvious to select the know T7G10 5’UTR and psbA 3’UTR as expression-control elements for predictable enhancement of plastid expression. Accordingly, claim 10 is obvious over Maliga and Lutz.
Claims 29, 45 and 53 are rejected under 35 U.S.C. §103 as being unpatentable over Maliga (US5877402A) as applied claim1, in view of Daniell (Henry Daniell, WO2001072959A2, Application: 2001-02-28, Publication: 2001-10-04).
Claim 1 as the teaching of Maliga is discussed above.
Claims 29 and 45 are interpreted as depend of claim 1.
Claim 29 recites the vector of claim 1, wherein the protein of interest is (b) expressed in an amount of about 0.1% to about 60% of total soluble protein (TSP) as species election.
Daniell teaches high-level accumulation of heterologous proteins in chloroplast, including about 7% of the total soluble protein of human somatotropin (page 111, line 6), 46.1% of the total soluble plant protein of Bt Cry2Aa2 operon (page 111, line 28). Daniell further teaches different construct modification to optimization of gene expression and protein production (page 114-123). It would have been obvious to optimize plastid expression within the claimed rage of about 0.1% to about 60% TSP using known expression control element. Claim 29 is obvious over Maliga and Daniell.
Claim 45 recites a plant plastid, plant cell or plant stably transformed with the transformation vector of claim 1, wherein: (c) the protein of interest is present in the plastid as a monomer, a dimer, a trimer or additional forms of multimers in amount of about 0.1% TSP to about 60% TSP as species election.
Daniell further teaches isolation of monomer fraction of HSA protein with Chromatographic techniques (page 89, line 24-27). Daniel further teaches, chloroplast expressed CTB protein shows CTB monomers predominantly, with some protein in the dimeric and trimeric form (page 58, line 1-14). It would have been obvious that a heterologous protein expressed in plastids may be present in monomeric, dimeric, trimeric, or multimeric forms depending on the biochemical nature of the protein. Claim 45 is obvious over Maliga and Daniell.
Claim 53 recites a progeny or seed of the plant of claim 45.
Daniell teaches stable plastid transformation of biologically active molecule (claim 9), wherein the biologically active molecule is HAS (claim 11), and “a stable transformed plant which comprises plastid stably transformed with the vector of claims 11, or the progeny or the seed thereof” (claim 12). Thus, progeny or seed of the transformed plant would have been taught.
Claim 53 is obvious over Maliga and Daniell.
Claim 16 is rejected under 35 U.S.C. §103 as being unpatentable over Maliga (US5877402A) as applied claim1, in view of Yarbakht (Melina Yarbakht et. al., Biotechnology and Applied Biochemistry (2015)62(1):55-63).
Claim 1 as the teaching of Maliga is discussed above.
Claim 16 is interpreted as depend of claim 1.
Claim 16 recites the vector of claim 1, wherein: (c) the protein of interest is Staphylococcus aureus Protein A as species election.
Maliga teaches a plastid/chloroplast transformation vector for expression of foreign proteins in plants (claim 1), and Maliga further teaches that plastids of higher plants are highly polyploid and present in many copies per cell, e.g., about 10,000 plastid genome copies per tobacco cell (column 43, line 40), making the plastid genome an attractive target for engineering because it provide readily obtainable high protein levels and permits high-level expression of recombinant proteins (column 57, line47-50).
Yarbakht teaches dicistronic expression of a human proinsulin-Protein A fusion in tobacco chloroplast, i.e., expression of a construct containing Staphylococcus aureus Protein A in the tobacco chloroplast genome( page 55, Abstract; page 56, left column, paragraph 3).
It would have been obvious to one of ordinary skill in the art at the time of the invention to select Staphylococcus aureus Protein A as the recited “protein of interest” in the plastid vector of Maliga because Yarbakht expressly teaches that Protein A can be expressed from the tobacco chloroplast, and Maliga teaches that plastid transformation in higher plants provide high-level expression due to the presence of multiple plastid genomes in each cell. Therefor, claim 16 is unpatentable over Maliga in view of Yarbakht.
Claim 18 is rejected under 35 U.S.C. §103 as being unpatentable over Maliga (US5877402A), in view of Yarbakht (2015) as applied claim 16, and further in view of Daniell (WO2001072959A2).
Claims 18 recites the vector of claim 16, wherein the plastid is from a dicot plant and wherein the dicot plant is a low-nicotine tobacco plant as species election.
For the same reason set forth as for claim 16, Daniell teaches plastid transformation in tobacco (claim 15), Daniell further teaches to use nicotine free edible tobacco to produce target protein (page 8, line 5-9). Such that the recited dicot low-nicotine plastid is taught.
Claim 18 is unpatentable over Maliga in view of Yarbakht and Daniell.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to YANXIN SHEN whose telephone number is (571)272-7538. The examiner can normally be reached Monday-Friday.
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/YANXIN SHEN/Examiner, Art Unit 1663
/WEIHUA FAN/Primary Examiner, Art Unit 1663
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