Compositions and methods for custom site-specific dna recombinases
AssigneeMonsanto Technology LLC
InventorsJonathan C. Lamb, Artem G. Evdokimov
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Abstract
The invention relates to biotechnology and provides novel compositions and methods for sequence-specific or sequence-directed transcription activator-like effector recombinases. The invention also provides novel plant transformation vectors and expression cassettes, which include novel combinations of chimeric recombinases with plant expression and transformation elements. Methods for gene-targeting, DNA sequence removal, and molecular breeding are also provided.
Claims53
A DNA construct encoding a chimeric recombinase polypeptide comprising a serine recombinase catalytic domain and a TALE DNA-binding domain comprising at least one repeat-variable di-residue (RVD) DNA-binding repeat domain.
An isolated nucleic acid sequence encoding a chimeric recombinase DNA-recombination target-sequence comprising, in a 5′ to 3′ orientation:
- a) a first TALE binding site;
- b) a TALER central sequence comprising, in a 5′ to 3′ orientation:
- i) an optional first spacer sequence;
- ii) a recombinase core sequence;
- and
- iii) an optional second spacer sequence;
- and
- c) a second TALE binding site.
A method of gene-targeting in a cell comprising introducing one or more chimeric recombinase polypeptides and one or more gene-targeting donor constructs comprising one or more TALER recombination sites into a cell, wherein a recombinase core sequence of one or more of said TALER recombination sites differs by 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 bp as compared to a recombinase core sequence in a genome of said cell.
Dependencies are inferred from the claim text.