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US 12,344,858 B2Granted
Targeted genome modification using circular single-stranded DNA
AssigneeFull Circles Therapeutics, Inc.
InventorsJin Hang Huh, Qun Shan
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Abstract
The present invention is directed to methods for generating one or more genetically modified cells by using a circular single stranded DNA (CiSSD) as a donor template and targeting genome modification. These methods include transferring one or more DNA polynucleotides into the cell for site-specific nuclease-mediated DNA repair and selecting one or more cells having the transferred DNA incorporated into the cell's genome.
Claims14
A method for generating a genetically modified cells, comprising:
- transferring a circular single stranded DNA (CiSSD) having a DNA insert, a 5′ homology arm, and a 3′ homology arm to a cell, wherein the 5′ homology arm and the 3′ homology arm are complementary to the polynucleotides in a target region of a genomic DNA in the cell;
- inducing a nucleotide break with Cas nuclease in the target region of genomic DNA in the cell;
- hybridizing the 5′ homology arm and the 3′ homology arm of the CiSSD with the complementary polynucleotides in the target region of the genomic DNA;
- and
- inserting the DNA insert into the target region of genomic DNA;
- whereby a genetically modified cell is generated, and the CiSSD has a higher integration efficiency than that using a linear single stranded DNA under the same condition.
3 more elements
Dependencies are inferred from the claim text.