Allele-specific genome editing using CRISPR-Cas9 is associated with loss of heterozygosity in diploid yeast.

ABSTRACT: Targeted DNA double-strand breaks (DSBs) with CRISPR-Cas9 have revolutionized genetic modification by enabling efficient genome editing in a broad range of eukaryotic systems. Accurate gene editing is possible with near-perfect efficiency in haploid or (predominantly) homozygous genomes. However, genomes exhibiting polyploidy and/or high degrees of heterozygosity are less amenable to genetic modification. Here, we report an up to 99-fold lower gene editing efficiency when editing individual heterozygous loci in the yeast genome. Moreover, Cas9-mediated introduction of a DSB resulted in large scale loss of heterozygosity affecting DNA regions up to 360 kb and up to 1700 heterozygous nucleotides, due to replacement of sequences on the targeted chromosome by corresponding sequences from its n

SUBMITTER: Gorter de Vries AR 

PROVIDER: S-EPMC6379674 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature