Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants.

ABSTRACT:

Background

Recent advances in clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing have led to the use of long single-stranded DNA (lssDNA) molecules for generating conditional mutations. However, there is still limited available data on the efficiency and reliability of this method.

Results

We generated conditional mouse alleles using lssDNA donor templates and performed extensive characterization of the resulting mutations. We observed that the use of lssDNA molecules as donors efficiently yielded founders bearing the conditional allele, with seven out of nine projects giving rise to modified alleles. However, rearranged alleles including nucleotide changes, indels, local rearrangements and additional integrat

SUBMITTER: Codner GF 

PROVIDER: S-EPMC6011369 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature