A simple and effective F0 knockout method for rapid screening of behaviour and other complex phenotypes

Kroll, François, Powell, Gareth T., Ghosh, Marcus, Gestri, Gaia, Antinucci, Paride, Hearn, Timothy J., Tunbak, Hande, Lim, Sumi, Dennis, Harvey W., Fernandez, Joseph M., Whitmore, David, Dreosti, Elena, Wilson, Stephen W., Hoffman, Ellen J. and Rihel, Jason (2021) A simple and effective F0 knockout method for rapid screening of behaviour and other complex phenotypes. eLife, 10. e59683. ISSN 2050-084X

Published Version
Available under the following license: Creative Commons Attribution.

Download (3MB) | Preview
Official URL: http://dx.doi.org/10.7554/elife.59683


Hundreds of human genes are associated with neurological diseases, but translation into tractable biological mechanisms is lagging. Larval zebrafish are an attractive model to investigate genetic contributions to neurological diseases. However, current CRISPR-Cas9 methods are difficult to apply to large genetic screens studying behavioural phenotypes. To facilitate rapid genetic screening, we developed a simple sequencing-free tool to validate gRNAs and a highly effective CRISPR-Cas9 method capable of converting &gt;90% of injected embryos directly into F0 biallelic knockouts. We demonstrate that F0 knockouts reliably recapitulate complex mutant phenotypes, such as altered molecular rhythms of the circadian clock, escape responses to irritants, and multi-parameter day-night locomotor behaviours. The technique is sufficiently robust to knockout multiple genes in the same animal, for example to create the transparent triple knockout <jats:italic>crystal</jats:italic> fish for imaging. Our F0 knockout method cuts the experimental time from gene to behavioural phenotype in zebrafish from months to one week.

Item Type: Journal Article
Faculty: Faculty of Science & Engineering
SWORD Depositor: Symplectic User
Depositing User: Symplectic User
Date Deposited: 10 Mar 2021 12:19
Last Modified: 31 Jan 2022 16:06
URI: https://arro.anglia.ac.uk/id/eprint/706403

Actions (login required)

Edit Item Edit Item