CRISPR-Cas9 Genome Editing in Nothobranchius furzeri for Gene Knockout and Knock-In

Itai Rozenberg, Eitan Moses, Itamar Harel*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

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.

Original languageAmerican English
Pages (from-to)90-99
Number of pages10
JournalCold Spring Harbor Protocols
Volume2023
Issue number2
DOIs
StatePublished - Feb 2023

Bibliographical note

Funding Information:
We thank the Harel laboratory for stimulating discussion and feedback. Supported by the Zuckerman Program (I.H.); Israel Science Foundation (ISF) 2178/19 (I.H.); Israel Ministry of Science 3-17631 (I.H.), 3-16872 (I.H.), and 3-16591 (E.M); the Moore Foundation GBMF9341 (I.H.) and BSF-NSF 2020611 (I.H.); and the Israel Ministry of Agriculture 12-16-0010 (I.H).

Publisher Copyright:
© 2023 Cold Spring Harbor Laboratory Press.

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