Melting dsDNA Donor Molecules Greatly Improves Precision Genome Editing in Caenorhabditis elegans.

ABSTRACT: CRISPR genome editing has revolutionized genetics in many organisms. In the nematode Caenorhabditis elegans, one injection into each of the two gonad arms of an adult hermaphrodite exposes hundreds of meiotic germ cells to editing mixtures, permitting the recovery of multiple indels or small precision edits from each successfully injected animal. Unfortunately, particularly for long insertions, editing efficiencies can vary widely, necessitating multiple injections, and often requiring coselection strategies. Here, we show that melting double-stranded DNA (dsDNA) donor molecules prior to injection increases the frequency of precise homology-directed repair (HDR) by several fold for longer edits. We describe troubleshooting strategies that enable consistently high editing efficiencie

SUBMITTER: Ghanta KS 

PROVIDER: S-EPMC7648581 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature