Improving one-step scarless genome editing in Drosophila melanogaster by combining ovo^D co-CRISPR selection with sgRNA target site masking.

ABSTRACT: The precise and rapid construction of alleles through CRISPR/Cas9-mediated genome engineering renders Drosophila melanogaster a powerful animal system for molecular structure-function analyses and human disease models. Application of the ovo^D co-selection method offers expedited generation and enrichment of scarlessly edited alleles without the need for linked transformation markers, which specifically in the case of exon editing can impact allele usability. However, we found that knockin procedures by homology-directed repair (HDR) under ovo^D co-selection resulted in low transformation efficiency. This is likely due to repeated rounds of Cas9 cleavage of HDR donor and/or engineered genomic locus DNA, as noted for other CRISPR/Cas9 editing strateg

SUBMITTER: Gotze KJ 

PROVIDER: S-EPMC8789338 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature