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-...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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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). |
format | Online Article Text |
id | pubmed-9617198 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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