CRISPR-associated transposases and uses thereof
AssigneeMonsanto Technology LLC
InventorsJaishree M. Chittoor, Ervin Nagy
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Abstract
Provided herein are systems, methods, and compositions for the modification of target DNA sequences. More particularly, systems, methods, and compositions for editing genomic DNA in eukaryotic cells with a CRISPR-associated transposase are provided. Also provided are vectors and vector systems which encode one or more CRISPR-associated transposases, as well as methods for the design and use of such vectors. Also provided are methods for identifying and validating novel CRISPR-associated transposases.
Claims20
A vector comprising a eukaryotic cell expressible promoter operably linked to a polynucleotide sequence encoding a CRISPR-associated enzyme, wherein the polynucleotide sequence has at least 85% identity to a sequence selected from the group consisting of SEQ ID NOs:
- 1-10, 12-97, 288-302, 304-594, 606-623, 2020-2069, 2075-2504, 2645-2679, 2700-2749, 2755-3184, and 3325-3359, and wherein the polynucleotide sequence encodes a protein having at least 85% identity to a sequence selected from the group consisting of SEQ ID NOs:
- 124-133, 135-220, and 277-283.
A method for sequence-specific modification of a target nucleic acid sequence in a eukaryotic cell, the method comprising providing to the eukaryotic cell:
- (a) a recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide sequence having at least 85% identity to a sequence selected from the group consisting of SEQ ID NOs:
- 1-10, 12-97, 288-302, 304-594, 606-623, 2020-2069, 2075-2504, 2645-2679, 2700-2749, 2755-3184, and 3325-3359, wherein the polynucleotide sequence encodes a protein having at least 85% identity to a sequence selected from the group consisting of SEQ ID NOs:
- 124-133, 135-220, and 277-283, and wherein the recombinant nucleic acid expresses an effective amount of a CRISPR-associated enzyme encoded by the polynucleotide sequence;
- and
- (b) a guide RNA capable of hybridizing with the target nucleic acid sequence, wherein the guide RNA forms a complex with the CRISPR-associated enzyme and the complex modifies the target nucleic acid sequence.
A recombinant nucleic acid comprising a heterologous promoter operably linked to a polynucleotide sequence encoding a protein with an amino acid sequence selected from the group consisting of SEQ ID NOs:
- 124-133, 135-220, and 277-283, wherein the polynucleotide sequence is codon optimized for expression in a plant cell.
Dependencies are inferred from the claim text.