San Diego, CA, United States
San Diego, CA, United States
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Provided herein include methods and compositions for making targeted changes to a DNA sequence. In various aspects and embodiments, methods and compositions for modifying a DNA sequence in a cell (such as a plant, bacterial, yeast, fungal, algal, or mammalian cell) are provided. In some aspects and embodiments the modification of DNA involves combining gene repair oligonucleotides with approaches that enhance the availability of components of the target cell gene repair mechanisms, such as a DNA cutter.


Provided herein include methods and compositions for effecting a targeted genetic change in DNA in a cell. Certain aspects and embodiments relate to improving the efficiency of the targeting of modifications to specific locations in genomic or other nucleotide sequences. As described herein, nucleic acids which direct specific changes to the genome may be combined with various approaches to enhance the availability of components of the natural repair systems present in the cells being targeted for modification.


Provided herein include methods and compositions for making targeted changes to a DNA sequence. In various aspects and embodiments, methods and compositions for modifying a DNA sequence in a cell (such as a plant, bacterial, yeast, fungal, algal, or mammalian cell) are provided. In some aspects and embodiments the modification of DNA involves combining gene repair oligonucleotides with approaches that enhance the availability of components of the target cell gene repair mechanisms, such as a DNA cutter.


Grant
Agency: Department of Agriculture | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 99.98K | Year: 2012

Solanaceous crops that include tomato and potato are multi-billion dollar crops annually and potato and tomato are the most widely grown specialty crops in the U.S. In general, introgression of genes from wild species that confer resistances to various stresses has been the foundation of most plant breeding through the 20th century to the present. This strategy has been particularly effective in crops such as tomato where the gene pool is narrow and genetic variation is found in related wild species. A major problem with interspecific transfer can be difficulty in introgressing the trait of interest into the target species. Fertilization barriers may demand special techniques, or linkage drag may bring in deleterious traits. Thus, new approaches for (i) direct identification of novel resistance traits in cultivated species and (ii) facile transfer into related species are needed. Approach (i) depends on multi-generation screening of ?mutator?tomato plants for novel resistance traits. Approach (ii) will use Cibus? Rapid Trait Development System (RTDS) to produce non-transgenic, specific mutations identified in approach (i), using chemically synthesized gene repair oligonucleotides (GRONs), in half the time of other breeding techniques. This technology harnesses the cell?s normal DNA-repair systems to correct and change specific targeted bases within the genome of a cell. The Gene Repair OligoNucleotide (GRON), a chemically synthesized oligonucleotide, is designed to create mismatched base-pairs compared to the target sequence within the host organism?s genome. The GRON hybridizes at the target region and the mismatched base-pairs work with the cell?s repair system at those sites to correct (replace, insert or delete) the designated base(s). Once the correction process is complete the GRON is degraded and the now-modified or repaired gene retains its normal pattern of expression and stability within the genome. Here we propose two linked proofs-of-concept for radically new ways to (i) select for novel crop variants resistant to different stresses and (ii) introduce novel resistance-conferring mutations into a related crop recalcitrant to breeding approaches. Approach (i) employs tomato as a prototype crop and Phytophthora infestans as a prototype pathogen and approach (ii) employs other solanaceous crops.


The invention provides to improved methods for the modification of genes in plant cells, and plants and seeds derived therefrom. More specifically, the invention relates to the increased efficiency of targeted gene mutation by combining gene repair oligonucleotides with approaches that enhance the availability of components of the target cell gene repair mechanisms.


Patent
Cibus Europe B.V. and Cibus Us Llc | Date: 2013-07-04

Provided are compositions and methods relating to gene and/or protein mutations in transgenic or non-transgenic plants. In certain embodiments, the disclosure relates to mutations in the protoporphyrinogen IX (PPX) gene. In some embodiments the disclosure relates to plants that are herbicide resistant.


The invention concerns the use of duplex oligonucleotides about 25 to 30 base pairs to introduce site specific genetic alterations in plant cells. The oligonucleotides can be delivered by mechanical (biolistic) systems or by electroporation of plant protoplasts. Thereafter plants having the genetic alteration can be generated form the altered cells. In specific embodiments the invention concerns the alteration in the gene that encode acid invertase, UDP-glucose pyrophosphorylase, polyphenol oxidase, O-methyl transferase, cinnamyl alcohol dehydrogenase, ACC synthase and ACC oxidase or etr-1 or homolog of etr-1, and plants having isolated point mutations in such genes.


Patent
Incima U.S. Llc and Cibus Us Llc | Date: 2012-09-17

The present invention relates to the production of a non-transgenic plant resistant or tolerant to a herbicide of the phosphonomethylglycine family, e.g., glyphosate. The present invention also relates to the use of a recombinagenic oligonucleobase to make a desired mutation in the chromosomal or episomal sequences of a plant in the gene encoding for 5-enol pyruvylshikimate-3-phosphate synthase (EPSPS). The mutated protein, which substantially maintains the catalytic activity of the wild-type protein, allows for increased resistance or tolerance of the plant to a herbicide of the phosphonomethylglycine family, and allows for the substantially normal growth or development of the plant, its organs, tissues or cells as compared to the wild-type plant irrespective of the presence or absence of the herbicide. Additionally the present invention relates to mutant E. coli cells that contain mutated EPSPS genes.


Patent
Cibus Us Llc and Cibus Europe B.V. | Date: 2014-03-14

The invention provides to improved methods for the modification of genes in plant cells, and plants and seeds derived therefrom. More specifically, the invention relates to the increased efficiency of targeted gene mutation by combining gene repair oligonucleotides with approaches that enhance the availability of components of the target cell gene repair mechanisms.


Patent
Cibus Us Llc and Cibus Europe B.V. | Date: 2014-03-14

Provided are compositions and methods relating to gene and/or protein mutations in plants. In certain embodiments, the disclosure relates to mutations in the allene oxide synthase 2 gene (i.e., AOS2). In some embodiments the disclosure relates to plants that are pathogen resistant.

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