Generation of knock-in lampreys by CRISPR-Cas9-mediated genome engineering.

ABSTRACT: The lamprey represents the oldest group of living vertebrates and has been a key organism in various research fields such as evolutionary developmental biology and neuroscience. However, no knock-in technique for this animal has been established yet, preventing application of advanced genetic techniques. Here, we report efficient generation of F₀ knock-in lampreys by CRISPR-Cas9-mediated genome editing. A donor plasmid containing a heat-shock promoter was co-injected with a short guide RNA (sgRNA) for genome digestion, a sgRNA for donor plasmid digestion, and Cas9 mRNA. Targeting different genetic loci, we succeeded in generating knock-in lampreys expressing photoconvertible protein Dendra2 as well as those expressing EGFP. With its simplicity, design flexibility, and high effic

SUBMITTER: Suzuki DG 

PROVIDER: S-EPMC8494898 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature