Methods and compositions for genome editing in plants
AssigneeNapigen, Inc.
InventorsHajime Sakai, Emil M. Orozco, Jr., Sachie Kimura, Elmer Heppard, Kamal Bhattarai, Mina Hamidi, Colleen McMichael, Cheryl S. Caster, Dawn Eigner, Debra J. Wilkinson
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Abstract
Various aspects of the present disclosure relate to methods and compositions for modulating or editing the genome of plants, including nuclear, plastid, and mitochondrial genomes. The disclosed methods employ genome editing tools, such as site-directed nucleases. Provided herein, in some aspects, are compositions (e.g., a plant) comprising the altered genome.
Claims20
A method for replacing an endogenous target polynucleotide present in a genome of a mitochondrion or a plastid of a cell with an internal polynucleotide, the method comprising:
- (a) introducing into the mitochondrion or the plastid of the cell a replacement nucleic acid comprising the internal polynucleotide,
- wherein the internal polynucleotide comprises a sequence heterologous to the genome of the mitochondrion or the plastid;
- (b) selecting an altered cell comprising at least one altered mitochondrion genome or at least one altered plastid genome, wherein the at least one altered mitochondrion genome or the at least one altered plastid genome comprises the internal polynucleotide;
- (c) introducing into the altered cell a polynucleotide encoding a site-directed nuclease (SDN), wherein the SDN binds to a target sequence present in the endogenous target polynucleotide;
- and
- (d) selecting a further altered cell, wherein at least about 50% of mitochondrial genomes of a plurality of mitochondrial genomes or at least about 50% of plastid genomes of a plurality of plastid genomes in the further altered cell comprise the internal polynucleotide.
A plant comprising a cell having an altered mitochondrion genome or an altered plastid genome, wherein the altered mitochondrion genome or the altered plastid genome comprises a heterologous polynucleotide comprising an internal polynucleotide in place of an endogenous polynucleotide,
- wherein the plant comprises a polynucleotide encoding a site-directed nuclease (SDN), wherein the SDN binds to a target sequence present in the endogenous polynucleotide, and
- wherein at least about 50% of mitochondrial genomes of a plurality of mitochondrial genomes or at least about 50% of the plastid genomes of a plurality of plastid genomes in the cell comprise the internal polynucleotide.
A polypeptide comprising a transcription activator-like (TAL) effector DNA-binding domain and a functional nuclease domain, wherein the functional nuclease domain comprises or consists of an amino acid sequence that is at least about 70% identical to an amino acid sequence of SEQ ID NO:
- 5 or SEQ ID NO:
- 60.
Dependencies are inferred from the claim text.