A simple and effective F0 knockout method for rapid screening of behaviour and other complex phenotypes
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- François Kroll
- Department of Cell and Developmental Biology, University College London
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- Gareth T Powell
- Department of Cell and Developmental Biology, University College London
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- Marcus Ghosh
- Department of Cell and Developmental Biology, University College London
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- Gaia Gestri
- Department of Cell and Developmental Biology, University College London
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- Paride Antinucci
- Department of Neuroscience, Physiology and Pharmacology, University College London
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- Timothy J Hearn
- Department of Cell and Developmental Biology, University College London
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- Hande Tunbak
- Wolfson Institute for Biomedical Research, University College London
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- Sumi Lim
- Department of Cell and Developmental Biology, University College London
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- Harvey W Dennis
- School of Biological Sciences, Faculty of Science, University of Bristol
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- Joseph M Fernandez
- Child Study Center, Yale School of Medicine
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- David Whitmore
- Department of Cell and Developmental Biology, University College London
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- Elena Dreosti
- Wolfson Institute for Biomedical Research, University College London
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- Stephen W Wilson
- Department of Cell and Developmental Biology, University College London
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- Ellen J Hoffman
- Child Study Center, Yale School of Medicine
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- Jason Rihel
- Department of Cell and Developmental Biology, University College London
書誌事項
- 公開日
- 2021-01-08
- 権利情報
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- http://creativecommons.org/licenses/by/4.0/
- http://creativecommons.org/licenses/by/4.0/
- http://creativecommons.org/licenses/by/4.0/
- DOI
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- 10.7554/elife.59683
- 公開者
- eLife Sciences Publications, Ltd
説明
<jats:p> 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 >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. </jats:p>
収録刊行物
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- eLife
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eLife 10 e59683-, 2021-01-08
eLife Sciences Publications, Ltd