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 .
Claim Status
The preliminary amendment filed 08/16/2024 is acknowledged. Claims 1-75 are cancelled. Claims 76-95 are new. Claims 76-95 are pending and under examination.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
Claims 90-93 are rejected under 35 U.S.C. 102a1 as being anticipated by Chan et al. (WO2020211782A1, IDS 08/16/24 Citation #6).
Claim 90 is drawn to a nucleic acid encoding a modified Cry3Aa protein and claims 91-93 are drawn to cells comprising a nucleic acid expressing a Cry3Aa protein.
Chan et al. teaches a modified Cry3Aa protein (Pos3Aa) in which select negatively charged amino acid residues are replaced with positively charged arginine or lysine to increase cellular uptake. More specifically, Chan et al. teaches that the positions N391, N395, E423, Q430, E432, E433, and E466, which are numbered according to SEQ ID NO: 1 (which corresponds to instant SEQ ID NO: 1), may be substituted for arginine or lysine [see claim 2] (instant claims 76, 77, 78, 79, 80). Given that Chan et al. teaches modifications to the claimed positions, the positions must necessarily fall within the solvent channel even if it is not expressly disclosed (instant claim 76). The resulting protein crystal has a solubility greater than 60% of wild-type Cry3Aa [see Fig. 11]. Chan et al. teaches incubating host cells under conditions that generate Cry3Aa protein crystals, isolating and solubilizing the protein crystals in an alkaline buffer, and subsequently recrystallizing the protein with a suitable buffer [see par. 74-78] (instant claims 94 and 95). Chan et al. teaches that an expression cassette comprising a promotor linked to a polynucleotide sequence encoding the modified protein can be expressed in Bacillus thuringiensis host cells [see claims 9-13] (instant claims 90-93).
Therefore, claims 90-93 are rejected under 35 U.S.C. 102a1.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
Claims 76-82, 87, and 90-93 are rejected under 35 U.S.C. 103 as being unpatentable over Chan et al. (WO2020211782A1, IDS 08/16/24 Citation #6), in view of Heater et al. (IDS 08/16/24 Citation #1), and in further view of Lilavivat et al.
Claims 76-82 are drawn to a Cry3Aa protein crystal, claim 87 is drawn to a plurality of Cry3Aa protein crystals.
The teachings of Chan et al. are discussed above.
Chan et al. does not teach or suggest encapsulation of a nucleic acid with the crystal.
Heater et al. teaches a Cry3Aa protein crystal (instant claim 76) having nanoporous solvent channels that can encapsulate a separate cargo molecule within the crystal [see abstract]. Heater et al. further teaches that cargo loading into Cry3Aa crystals is affected by electrostatic interactions between the cargo and the Cry3Aa crystal solvent channels and demonstrates that altering the charge of the channel changes the amount of cargo incorporated into the crystal [see p. 9880, col. 2, par. 3 and 9881, col. 1, par. 1]. Heater et al. teaches that the Cry3Aa protein is natively expressed in Bacillus thuringiensis [see abstract].
Lilavivat et al. teaches that increasing the positive charge of the interior of a protein container results in uptake and enrichment of negatively charged RNA (instant claim 82) into the container during assembly [see abstract] (instant claim 76).
It would have been obvious to combine these teachings to generate the claimed protein. Heater et al. establishes that Cry3Aa crystals can encapsulate cargo and teaches that electrostatic interactions, including the charge of the crystal solvent channel, affect cargo loading. Chan et al. teaches a specific method of modifying the charge of the Cry3Aa by replacing specific residues with positively charged lysine or arginine and further teaches that such crystals retain their ability to form crystals and exhibit enhanced uptake by target cells. Lilavivat et al. teaches that a positively charged protein interior can be used to encapsulate negatively charged RNA through electrostatic charge complementarity. Thus, a person having ordinary skill in the art would have understood that negatively charged nucleic acids could be incorporated into molecular containers using charge complementarity, that Cry3Aa proteins form crystals that could function as molecular containers for cargo, and that the electrostatic environment of the Cry3Aa crystal could be modified to enhance binding via interactions with the cargo (instant claim 76) while also increasing protein uptake in the target cells. There would have been a reasonable expectation of success because each component of the proposed combination relies on mechanisms and modifications that had already been demonstrated in the art to be successful, with the general principle of electrostatic interactions being a mechanism that is not limited to any specific scaffold protein.
Similarly, it would have been obvious to generate the protein by expressing it in a host cell, such as Bacillus thuringiensis, via an expression cassette linked to a promotor because the art teaches that the protein properly forms under these conditions and that Cry3Aa is natively expressed in Bacillus thuringiensis (instant claims 90-93). It would have been obvious to generate a plurality of the modified Cry3Aa protein as the art teaches that the protein is useful for targeting tissues with therapeutics and efficacious administration would require more than one of the protein to be present (instant claim 87).
Regarding claim 81, the limitation of “wherein the solubility of the Cry3Aa protein crystal is greater than 60% of the solubility of a wild-type Cry3A protein crystal, wherein solubility is determined in a 15 mL solubilization reaction at pH 11 and a crystal concentration of 1 mg/ml” is interpreted as an inherent property of a structure that meets all requirements of the claimed composition.
Therefore, claims 76-82, 87, and 90-93 are rejected under 35 U.S.C. 103.
Claims 76, 82-86, 88, and 89 are rejected under 35 U.S.C. 103 as being unpatentable over Chan et al. (WO2020211782A1, IDS 08/16/24 Citation #6), in view of Heater et al. (IDS 08/16/24 Citation #1), and in further view of Lilavivat et al. and Ochoa-Campuzano et al.
Claim 76 and 82-86 are drawn to a Cry3Aa protein crystal and claims 88 and 89 are drawn to a method of using a Cry3Aa protein crystal.
The disclosure of Chan et al., Heater et al., and Lilavivat et al. are discussed above.
The combination of these references does not teach or suggest the inclusion of an RNAi associated molecule or using the protein to control a pest population.
Ochoa-Campuzano et al. teaches RNAi-mediated silencing of the prohibitin-1 gene in Colorado potato beetle larvae via double stranded RNA (dsRNA) [see p. 302, col. 1, par. 1-2] produces deleterious effects in the pest (instant claims 83, 85, and 86) [see abstract]. Ochoa-Campuzano et al. teaches that RNAi can be co-administered with Cry3Aa to achieve highly effective pest mortality and identifies the combination as an effective strategy for crop protection [see abstract].
It would have been obvious to combine these teachings and select an RNAi associated nucleic acid as the negatively charged biomolecule of the modified Cry3Aa crystal discussed above because Ochoa-Campuzano et al. teaches that simultaneous use of these two compositions is an effective means of controlling crop pests and would therefore provide an additional mechanism for suppressing said pests (instant claim 82). Further, it would have been obvious that the dsRNA method taught by Ochoa-Campuzano et al. could be modified to instead use siRNA, shRNA, miRNA, or any similar cargo as all such molecules were well established components to be used in RNAi, and a skilled artisan could have easily selected an appropriate RNAi format based on the target organism, gene, and mechanism of silencing (instant claim 84). There would have been a reasonable expectation of success as the efficacy of each component was individually known and it would have been expected that this nucleic acid would be encapsulated no differently than any other.
Similarly, it would have been obvious to deliver an effective amount of the composition to a pest such as the Colorado potato beetle larvae because Ocoha-Campuzano et al. teaches that both the RNAi method as well as Cry3Aa are both useful methods for controlling said pests (instant claim 88). It would have been obvious to apply the composition to a crop plant because doing so is the conventional means of implementing the pesticide strategy (instant claim 89).
Therefore, claims 76, 82-86, 88, and 89 are rejected under 35 U.S.C. 103.
Claims 94 and 95 are rejected under 35 U.S.C. 103 as being unpatentable over Chan et al. (WO2020211782A1, IDS 08/16/24 Citation #6), in view of Heater et al. (IDS 08/16/24 Citation #1), and in further view of Lilavivat et al. and Lucignano et al.
Claims 94 and 95 are drawn to a method of making a Cry3Aa protein crystal.
The disclosure of Chan et al., Heater et al., and Lilavivat et al. are discussed above.
The combination of these references does not teach or suggest the specific method of making the claimed protein.
Lucignano et al. teaches that ferritin nanocages can be disassembled and reassembled in vitro by modifying the pH of the solution, which allows loading of cargo molecules into the cages. Lucignano et al. teaches that pH and temperature are optimizable experimental conditions depending on the protein used and the cargo to be encapsulated [see abstract].
It would have been obvious to combine these teachings and utilize the cargo loading strategy of Lucignano et al. in the Cry3Aa system discussed above because Chan et al. teaches that Cry crystals can be solubilized and renatured and Lucignano et al. teaches that disassembly and reassembly in the presence of cargo is an effective method of loading said cargo into the carrier (instant claim 94). There would have been a reasonable expectation of success as the Cry3Aa protein demonstrated the necessary characteristics to employ said method, that being the ability to solubilize and recrystallize in the presence of cargo.
Furthermore, it would have been obvious to generate the method of instant claim 95 as Chan et al. teaches incubating host cells under conditions that generate Cry3Aa protein crystals, isolating and solubilizing the protein crystals in an alkaline buffer, and subsequently recrystallizing the protein with a suitable buffer, while Lucignano et al. teaches that pH adjustment is used to control reversible protein disassembly and reassembly and that cargo can be introduced during the reassembly process. A person having ordinary skill in the art would have therefore understood that introducing the biomolecule in a buffer having a pH selected to move the crystal towards conditions favorable for reassembly to be a matter of routine optimization well within the skill of the ordinary artisan (instant claim 95).
Therefore, claims 94 and 95 are rejected under 35 U.S.C. 103.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Tirone D Johnson whose telephone number is (571)272-1256. The examiner can normally be reached M-F, 9-5 ET.
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/TIRONE D. JOHNSON/ Examiner, Art Unit 1675
/JEFFREY STUCKER/ Supervisory Patent Examiner, Art Unit 1675
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