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 .
Status of Claims
The amendments received on 03/18/2026 have been entered. Claims 17-18, 20-21, 23-25, and 30-31 are pending.
Claims 20, 24, and 30-31 remain withdrawn.
Claims 17-18 have been amended.
Claims 17-18, 21, 23, and 25 are examined in this Office Action.
The text of those sections of Title 35, U.S. Code, not included in this action, can be found in a prior Office action.
Information Disclosure Statement
Initialed and dated copy of Applicant’s information disclosure statement (IDS) filed on
03/18/2026 is attached to the instant Office Action. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Claim Rejections - 35 USC § 103
Claims 17, 18, and 25 are rejected under 35 U.S.C. 103 as being unpatentable over DAVIS (Davis et al., Pub. No.: US 2019/0292217 A1, Pub. Date: Sep. 26, 2019; see List of references cited by examiner dated 05/12/2025) in view of JARDINAUD (Jardinaud et al., International Publication Number: WO 99/02687, International Publication Date: 21 January 1999) in further view of BANSAL (Bansal et al., 2012, Agricultural Research, Vol. 1, pp. 53-66). This is a modified rejection necessitated by amendment.
Claim 17 recites “[a] method of producing a myoglobin protein in a transgenic plant, wherein the method comprises:
(a) growing the transgenic plant, wherein the transgenic plant is a Lactuca species and comprises at least one chloroplast with one or more recombinant nucleic acid sequences expressing a bovine myoglobin gene encoding the myoglobin protein, wherein the transgenic plant comprises at least about 2,000 copies of the one or more recombinant nucleic acid sequences integrated into the chloroplast DNA of the transgenic plant, and
(b) isolating the myoglobin protein from the transgenic plant.
DAVIS teaches and claims methods and materials for making transgenic plants, transgenic plant cells, and transgenic seeds in which heme biosynthesis is specifically upregulated (Davis, page 1, 0004); a transgenic plant comprising at least one recombinant nucleic acid, wherein the recombinant nucleic acid comprises a first promoter operably linked to a nucleic acid encoding a heme-containing polypeptide (Davis, claim 1) (i.e., method of producing a myoglobin protein in a transgenic plant).
DAVIS teaches the heme-containing polypeptide can be a bovine myoglobin (i.e., one or more recombinant nucleic acid sequences expressing a bovine myoglobin gene encoding the myoglobin protein) (Davis, page 3, 0033).
DAVIS teaches and claims growing a plant cell comprising at least one recombinant nucleic acid (i.e., growing the transgenic plant) (Davis, page 2, 0017; claim 52).
DAVIS teaches the nucleic acid construct further includes a targeting sequence that can be used to direct the heme polypeptide and/or heme biosynthesis polypeptide to one of several different intracellular compartments, including, for example, plastids (such as chloroplasts) (i.e., wherein the one or more recombinant nucleic acid sequences is integrated into the chloroplast DNA of the transgenic plant) (Davis, page 6, 0050).
DAVIS teaches the transgenic plants or plant cells can be grown in a manner suitable for the species under consideration, and then the heme-loaded polypeptide can be isolated (i.e., isolating the myoglobin protein from the transgenic plant) (Davis, page 7, 0060). To isolate the heme-loaded heme protein from the transgenic plants, cells, or seeds, the plant material can be processed using any appropriate measure (Davis, page 7, 0063).
DAVIS does not explicitly teach wherein the transgenic plant is a Lactuca species or wherein the transgenic plant comprises at least about 2,000 copies of the one or more recombinant nucleic acid sequences.
However, JARDINAUD teaches that iron deficiency is the most prevalent nutritional disorder worldwide. Individuals from socio-economically disadvantaged groups, ethnic groups (Aborigines, Asian migrants), vegetarians, and athletes are considered to be at particular risk from iron deficiency. Other groups at high risk are adolescents with increased requirements associated with growth and puberty, particularly females with the commencement of menstruation (Jardinaud, Background to The Invention, page 1, lines 25-26, and page 2, lines 4-8).
JARDINAUD teaches that nutritional iron problems associated with vegetarian diets are particularly prevalent in developing countries where major reliance is placed on staple cereals such as rice and wheat, and where intake of animal or fish sources of heme protein are either culturally unacceptable or impractical due to shortage or unreliability of supply, or poverty (Jardinaud, Background to The Invention, page 2, lines 10-13).
JARDINAUD sought to increase the bioavailability to humans and other animals, of iron in non-animal foodstuffs, such as plants, by introducing thereto an iron-binding protein molecule. In particular, JARDINAUD discovered that the bioavailability of iron may be increased by introducing a genetic sequence which encodes a heme protein into a plant cell and expressing said genetic sequence therein (Jardinaud, Background to The Invention, page 3, lines 15-20).
JARDINAUD teaches that the iron-binding protein is a heme protein, more preferably a hemoglobin, myoglobin or ferritin polypeptide or a homologue, analogue or derivative thereof (Jardinaud, page 13, lines 5-7).
JARDINAUD teaches plants which may be employed in practicing the present invention include all edible plants such as lettuce (e.g., Lactuca sativa) (i.e., wherein the transgenic plant is a Lactuca species) (Jardinaud, page 33, lines 6-19).
Neither DAVIS nor JARDINAUD explicitly teaches wherein the transgenic plant comprises at least about 2,000 copies of the one or more recombinant nucleic acid sequences.
BANSAL teaches that plastid transformation in higher plants has been established in the recent past to engineer several agronomic traits including herbicide resistance, insect and pathogen resistance, abiotic stress tolerance, increased photosynthesis and also production of edible crops engineered to produce ‘biopharmaceuticals’. Research efforts led to the development of this technology in several crops plants, like potato, tomato, brinjal, rice, wheat, oilseed rape, soybean, and in lettuce. The progress reported thus far has helped raise hopes for generating transplastomic crops in the near future with engineered agronomic traits (Bansal, page 54, left column, last paragraph).
BANSAL further teaches that the copy number of plastid genome per plant cell is very high; each chloroplast consists of 50–100 copies of plastid genome and each cell consists of more than hundreds of chloroplasts making the copy number of ~10,000 per cell. This feature of high copy number of genomes is exploited in genetic engineering of plastids for over expression of transgenes, thereby allowing the recombinant proteins to accumulate at high concentrations of over 10% of the total soluble proteins (i.e., wherein the transgenic plant comprises at least about 2,000 copies of the one or more recombinant nucleic acid sequences, instant claim 17; wherein the transgenic plant comprises at least about 5,000 copies of the one or more recombinant nucleic acid sequences, instant claim 18) (Bansal, page 54, right column, last paragraph to page 55, left column, first paragraph).
At the time the instant application was filed, it would have been obvious and within the scope of one having ordinary skill in the art to use the method of producing a myoglobin protein in a transgenic plant as taught by Davis, in a plant of the Lactuca species as taught by Jardinaud, comprising at least about 2,000 copies of the one or more recombinant nucleic acid sequences as taught by Bansal, as required by the claims. One would have been motivated to use the method of producing a myoglobin protein in a transgenic plant as taught by Davis, in a plant of the Lactuca species as taught by Jardinaud, knowing that a Lactuca species is a suitable host for producing a large amount of myoglobin protein in a transgenic plant as taught by Jardinaud and Bansal. Thus, one of ordinary skill in the art would have a high expectation of success by following the teachings of Davis, Jardinaud, and Bansal since the method of producing a myoglobin protein in a transgenic plant is well-known in the art and has been employed to increase the accumulation of a target protein for decades. Additionally, it is well-known in the art that routine optimization of protein production would preferably require multiple copies of the gene of interest to be integrated into the chloroplast DNA of the plant; the number of copies being a mere design choice and routine optimization in the art.
In regard to claim 18, see claim 17 rejection above.
In regard to claim 25, DAVIS teaches and claims a transgenic plant comprising at least one recombinant nucleic acid, wherein the recombinant nucleic acid comprises a first promoter operably linked to a nucleic acid encoding a heme-containing polypeptide (i.e., wherein the myoglobin gene is operably linked to at least one promoter) (Davis, page 1, 0005; claim 1).
Claims 21 and 23 are rejected under 35 U.S.C. 103 as being unpatentable over DAVIS (Davis et al., Pub. No.: US 2019/0292217 A1, Pub. Date: Sep. 26, 2019; see List of references cited by examiner dated 05/12/2025), JARDINAUD (Jardinaud et al., International Publication Number: WO 99/02687, International Publication Date: 21 January 1999) and BANSAL (Bansal et al., 2012, Agricultural Research, Vol. 1, pp. 53-66) as applied to claims 17, 18, and 25 above, in further view of TRAN (Tran et al., Pub. No.: US 2020/0332249 A1, Pub. Date: Oct. 22, 2020; see List of references cited by examiner dated 12/16/2024). This is a modified rejection necessitated by amendment.
Claim 21 recites “[t]he method of claim 17, wherein the transgenic plant is: (a) a stable, homoplasmic transformant; or (b) a stable heteroplasmic transformant”.
DAVIS, JARDINAUD, and BANSAL teach the method of claim 17.
DAVIS, JARDINAUD, and BANSAL do not teach wherein the transgenic plant is: (a) a stable, homoplasmic transformant; or (b) a stable heteroplasmic transformant”.
TRAN teaches that the expression of the iron-complexed protein can be accomplished by inserting a nucleic acid molecule (gene) encoding the protein into the chloroplast and/or nuclear genome of a microalgae. The modified strain of microalgae can be made homoplasmic to ensure that the polynucleotide will be stably maintained in the chloroplast genome of all descendants. A microalga is homoplasmic for a gene when the inserted gene is present in all copies of the chloroplast genome, for example. It is apparent to one of skill in the art that a chloroplast may contain multiple copies of its genome, and therefore, the term “homoplasmic” or “homoplasmy” refers to the state where all copies of a particular locus of interest are substantially identical. Plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% or more of the total soluble plant protein (page 10, paragraph 0073).
TRAN further teaches the chloroplast genome of two Chlamydomonas strains, THN1 and THN6, were transformed with a bovine myoglobin gene. Once homoplasmic, the strains of Chlamydomonas reinhardtii containing the bovine myoglobin gene were grown in a flask in the dark and light. THN6+myoglobin grown in the dark is shown to accumulate myoglobin (i.e., the transgenic plant is a stable homoplasmic transformant) (Examples, page 15, paragraph 0106).
At the time the instant application was filed, it would have been obvious and within the scope of one having ordinary skill in the art to strive for homoplasmy as taught by Tran, in the transgenic Lactuca plant as taught by Davis, Jardinaud, and BANSAL, as required by the claims. One would have been motivated to strive for homoplasmy as taught by Tran, in the transgenic Lactuca plant as taught by Davis, Jardinaud, and BANSAL, to ensure that the polynucleotide will be stably maintained in the chloroplast genome of all descendants. Thus, one of ordinary skill in the art would have a high expectation of success by following the teachings of Davis, Jardinaud, Bansal, and Tran, since homoplasmy is well-known in the art and has been employed to stably maintain a polynucleotide in the chloroplast genome of all descendants for decades.
In regard to claim 23, TRAN teaches SEQ ID NO: 13 (bovine myoglobin amino acid sequence) (i.e., bovine myoglobin) which shares 100% identity to instant sequence SEQ ID NO: 1 (myoglobin protein selected from SEQ ID NOs: 1-3 and 5-6) (see alignment below).
SEQUENCE ALIGNMENT BETWEEN TRAN SEQ ID NO:13 AND INSTANT SEQUENCE SEQ ID NO: 1
Qy 1 MGLSDGEWQLVLNAWGKVEADVAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASE 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MGLSDGEWQLVLNAWGKVEADVAGHGQEVLIRLFTGHPETLEKFDKFKHLKTEAEMKASE 60
Qy 61 DLKKHGNTVLTALGGILKKKGHHEAEVKHLAESHANKHKIPVKYLEFISDAIIHVLHAKH 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 DLKKHGNTVLTALGGILKKKGHHEAEVKHLAESHANKHKIPVKYLEFISDAIIHVLHAKH 120
Qy 121 PSDFGADAQAAMSKALELFRNDMAAQYKVLGFHG 154
||||||||||||||||||||||||||||||||||
Db 121 PSDFGADAQAAMSKALELFRNDMAAQYKVLGFHG 154
Response to Applicant’s Arguments
Applicant's arguments filed 03/18/2026 have been fully considered but they are not persuasive. Initially, it is noted that this is a modified rejection necessitated by amendment.
Applicant argues in part (A) that the combination of Davis and Jardinaud fails to teach or suggest all claimed elements (Remarks, pages 4-6).
The Examiner respectfully disagrees. Applicant has added the limitation wherein the “transgenic plant comprises at least about 2,000 copies of the” one or more recombinant nucleic acid sequences. The instant Specification describes that the stable chloroplast transformation in plants provides for commercial scale manufacturing of myoglobin meat protein in transgenic plants. There are unique advantages of chloroplast transformation technologies where the recombinant genes of interest are integrated into a targeted site of the chloroplast genome by homologous recombination. For example, non-limiting examples of chloroplast transformation can include: a) higher expression of foreign genes because of multiple copies (1,000-50,000 copies) of the genes due to the multi-copy of chloroplast DNA (100-250 copies) per chloroplast and multi-copy of chloroplasts in the cells (instant Specification, page 2, paragraph 0007). The instant Specification is silent as to the criticality of 2,000 copies of the recombinant nucleic acid sequence.
Although Davis and Jardinaud do not explicitly teach the “transgenic plant comprises at least about 2,000 copies of the” one or more recombinant nucleic acid sequences, the introduction of Bansal overcomes this deficiency with the teaching that the copy number of plastid genome per plant cell is very high; each chloroplast consists of 50–100 copies of plastid genome and each cell consists of more than hundreds of chloroplasts making the copy number of ~10,000 per cell. This feature of high copy number of genomes is exploited in genetic engineering of plastids for over expression of transgenes, thereby allowing the recombinant proteins to accumulate at high concentrations of over 10% of the total soluble proteins (see claim 17 rejection above). Additionally, Tran reinforces the teachings of Bansal in stating that plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% or more of the total soluble plant protein (see claim 21 rejection above).
Applicant argues in part (B) that the combination of Davis, Jardinaud, and Tran does not provide a reasonable expectation of success to arrive at the claimed invention (Remarks, pages 7-9).
The Examiner respectfully disagrees. Although the combination of Davis, Jardinaud, and Tran does not explicitly provide the specific copy number of 2,000, Tran does teach that plastid expression, in which genes are inserted by homologous recombination into all of the several thousand copies of the circular plastid genome present in each plant cell, takes advantage of the enormous copy number advantage over nuclear-expressed genes to permit expression levels that can readily exceed 10% or more of the total soluble plant protein (see claim 21 rejection above). This teaching from Tran would lead one of ordinary skill in the art to expect a significant increase in protein production with the enormous copy number advantage of the plastid genome.
Applicant additionally argues in part (B) that obviousness can be overcome by a showing of unexpected results. The instant Specification does not disclose any quantified results of myoglobin accumulation in the transformed soybean, tobacco, or lettuce plants, only that myoglobin was successfully accumulated, via Western Blot results. In the Remarks (dated 03/18/2026, pages 9-10), Applicant has introduced a figure and measurements that are not included in the instant Specification. Applicant states “an observed myoglobin accumulation of approximately 1.5% of total soluble protein (TSP), corresponding to about 0.814mg/g dry weight (DW)”, which is not disclosed in the instant Specification. Applicant is invited to submit a Declaration with supporting evidence of the allegedly unexpected results. Submission of evidence solely based on Attorney’s arguments is not sufficient.
Summary
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Correspondence
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHRISTINA MEADOWS whose telephone number is (703)756-1430. The examiner can normally be reached Monday - Friday 9:00 am - 5:00 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Amjad Abraham can be reached at 571-270-7058. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
CHRISTINA MEADOWS
Examiner
Art Unit 1663
/CHRISTINA L MEADOWS/Examiner, Art Unit 1663
/Amjad Abraham/SPE, Art Unit 1663
Prosecution Insights