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US 12,070,022 B2Granted
Methods for making genetic edits
AssigneeRecombinetics, Inc.
InventorsScott C. Fahrenkrug, Daniel F. Carlson
Life of this patent
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Abstract
The present disclosure relates to methods for making genetic edits in vitro in a non-human vertebrate cell or embryo at a plurality of target chromosomal DNA sites. Methods for making a non-human animal having multiplex genetic edits at a plurality of target chromosomal DNA sites and making a non-human vertebrate animal chimeric for host cells and donor cells are also considered.
Claims13
A method of making gene edits in vitro in a non-human vertebrate cell at two or more gene loci, the method comprising a simultaneous step of introducing into a non-human vertebrate cell:
- a first targeted CRISPR endonuclease system directed to a first target chromosomal DNA site in a genome of the non-human vertebrate cell,
- a first homology directed repair template homologous to a single gene locus in the first target chromosomal DNA site,
- a second targeted CRISPR endonuclease system directed to a second target chromosomal DNA site in the genome of the non-human vertebrate cell, and
- a second homology directed repair template homologous to a single gene locus in the second target chromosomal DNA site,
- wherein the first homology directed repair template sequence independently replaces a native chromosomal DNA sequence at the single gene locus in the first target chromosomal DNA site in the genome of the non-human vertebrate cell and the second homology directed repair template sequence independently replaces a native chromosomal DNA sequence at the single gene locus in the second target chromosomal DNA site in the genome of the non-human vertebrate cell;
- wherein the first target chromosomal DNA site is in a first gene and the second target chromosomal DNA site is in a second gene;
- thereby making a non-human vertebrate cell comprising gene edits at two or more gene loci.
4 more elements
Dependencies are inferred from the claim text.