Targeted genomic modification with partially single-stranded donor molecules

摘要

Disclosed herein are donor molecules comprising single-stranded complementary regions flanking one or more sequences of interest. The donor molecules and/or compositions comprising these molecules can be used in methods for targeted integration of an exogenous sequence into a specified region of interest in the genome of a cell.

主权项

1. A mammalian or plant cell comprising:
a genome comprising a target site of interest; a non-naturally occurring zinc finger nuclease that is capable of generating a double-stranded break with single-stranded overhangs in the genome at the target site of interest; and an exogenous linear donor nucleic acid molecule comprising a double-stranded exogenous sequence of interest having first and second ends and first and second single-stranded sequences of 4 to 10 nucleotides at the first and second ends of the double-stranded sequence, wherein the first and second single-stranded sequences are 100% complementary to the single-stranded overhangs in the genome at the target site of interest such that the exogenous linear donor sequence is capable of being directly integrated at the target site of interest by annealing of the complementary first and second single-stranded sequences and non-homologous end joining (NHEJ)-mediated targeted integration.