CRISPR–Cas9 Genome Editing in Nothobranchiusfurzerifor Gene Knockout and Knock-In

Autor: Rozenberg, Itai, Moses, Eitan, Harel, Itamar
Zdroj: Cold Spring Harbor Protocols; 2023, Vol. 2023 Issue: 2 ppdb.prot10-pdb.prot10, 1p
Abstrakt: The African turquoise killifish Nothobranchius furzerihas recently gained interest as an emerging vertebrate model system for the study of aging, owing to its naturally short life span and generation time. Here, we provide a step-by-step guide for effective genome engineering using the CRISPR–Cas9 system to generate loss-of-function (i.e., knockout) alleles and for precise editing (i.e., knock-in) of short sequences into the genome. Using this approach, a new stable line can be created within several months. The killifish's tough chorion, rapid growth, and short life span are considered in this protocol and account for the key deviations from similar protocols in other fish models.
Databáze: Supplemental Index