CRISPR-Cas9-induced gene knockout in zebrafish

The application of CRISPR has greatly facilitated genotype-phenotype studies of human disease models. In this protocol, we describe CRISPR-Cas9-induced gene knockout in zebrafish, utilizing purified Cas9 protein and in vitro-transcribed sgRNA. This protocol targets the PHLPP1 gene in an Indian wild-...

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Autores principales: Medishetti, Raghavender, Balamurugan, Keerthana, Yadavalli, Krishnaveni, Rani, Rita, Sevilimedu, Aarti, Challa, Anil Kumar, Parsa, Kishore, Chatti, Kiranam
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9617198/
https://www.ncbi.nlm.nih.gov/pubmed/36317180
http://dx.doi.org/10.1016/j.xpro.2022.101779
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author Medishetti, Raghavender
Balamurugan, Keerthana
Yadavalli, Krishnaveni
Rani, Rita
Sevilimedu, Aarti
Challa, Anil Kumar
Parsa, Kishore
Chatti, Kiranam
author_facet Medishetti, Raghavender
Balamurugan, Keerthana
Yadavalli, Krishnaveni
Rani, Rita
Sevilimedu, Aarti
Challa, Anil Kumar
Parsa, Kishore
Chatti, Kiranam
author_sort Medishetti, Raghavender
collection PubMed
description The application of CRISPR has greatly facilitated genotype-phenotype studies of human disease models. In this protocol, we describe CRISPR-Cas9-induced gene knockout in zebrafish, utilizing purified Cas9 protein and in vitro-transcribed sgRNA. This protocol targets the PHLPP1 gene in an Indian wild-caught strain, but is broadly applicable. Major factors influencing protocol success include zebrafish health and fecundity, sgRNA efficiency and specificity, germline transmission, and mutant viability. For complete details on the use and execution of this protocol, please refer to Balamurugan et al. (2022).
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spelling pubmed-96171982022-10-30 CRISPR-Cas9-induced gene knockout in zebrafish Medishetti, Raghavender Balamurugan, Keerthana Yadavalli, Krishnaveni Rani, Rita Sevilimedu, Aarti Challa, Anil Kumar Parsa, Kishore Chatti, Kiranam STAR Protoc Protocol The application of CRISPR has greatly facilitated genotype-phenotype studies of human disease models. In this protocol, we describe CRISPR-Cas9-induced gene knockout in zebrafish, utilizing purified Cas9 protein and in vitro-transcribed sgRNA. This protocol targets the PHLPP1 gene in an Indian wild-caught strain, but is broadly applicable. Major factors influencing protocol success include zebrafish health and fecundity, sgRNA efficiency and specificity, germline transmission, and mutant viability. For complete details on the use and execution of this protocol, please refer to Balamurugan et al. (2022). Elsevier 2022-10-26 /pmc/articles/PMC9617198/ /pubmed/36317180 http://dx.doi.org/10.1016/j.xpro.2022.101779 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Medishetti, Raghavender
Balamurugan, Keerthana
Yadavalli, Krishnaveni
Rani, Rita
Sevilimedu, Aarti
Challa, Anil Kumar
Parsa, Kishore
Chatti, Kiranam
CRISPR-Cas9-induced gene knockout in zebrafish
title CRISPR-Cas9-induced gene knockout in zebrafish
title_full CRISPR-Cas9-induced gene knockout in zebrafish
title_fullStr CRISPR-Cas9-induced gene knockout in zebrafish
title_full_unstemmed CRISPR-Cas9-induced gene knockout in zebrafish
title_short CRISPR-Cas9-induced gene knockout in zebrafish
title_sort crispr-cas9-induced gene knockout in zebrafish
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9617198/
https://www.ncbi.nlm.nih.gov/pubmed/36317180
http://dx.doi.org/10.1016/j.xpro.2022.101779
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