TY - JOUR
T1 - CRISPR-Cas9 Genome Editing in Nothobranchius furzeri for Gene Knockout and Knock-In
AU - Rozenberg, Itai
AU - Moses, Eitan
AU - Harel, Itamar
N1 - Publisher Copyright:
© 2023 Cold Spring Harbor Laboratory Press.
PY - 2023/2
Y1 - 2023/2
N2 - The African turquoise killifish Nothobranchius furzeri has 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.
AB - The African turquoise killifish Nothobranchius furzeri has 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.
UR - http://www.scopus.com/inward/record.url?scp=85147317795&partnerID=8YFLogxK
U2 - 10.1101/pdb.prot107742
DO - 10.1101/pdb.prot107742
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C2 - 36223984
AN - SCOPUS:85147317795
SN - 1940-3402
VL - 2023
SP - 90
EP - 99
JO - Cold Spring Harbor Protocols
JF - Cold Spring Harbor Protocols
IS - 2
ER -