Cargando…
Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos
CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA kn...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715325/ https://www.ncbi.nlm.nih.gov/pubmed/35005640 http://dx.doi.org/10.1016/j.xpro.2021.101058 |
_version_ | 1784624106450190336 |
---|---|
author | Hernandez-Huertas, Luis Kushawah, Gopal Diaz-Moscoso, Alejandro Tomas-Gallardo, Laura Moreno-Sanchez, Ismael da Silva Pescador, Gabriel Bazzini, Ariel A. Moreno-Mateos, Miguel A. |
author_facet | Hernandez-Huertas, Luis Kushawah, Gopal Diaz-Moscoso, Alejandro Tomas-Gallardo, Laura Moreno-Sanchez, Ismael da Silva Pescador, Gabriel Bazzini, Ariel A. Moreno-Mateos, Miguel A. |
author_sort | Hernandez-Huertas, Luis |
collection | PubMed |
description | CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo optimized CRISPR-RfxCas13d (CasRx) system. Although the described protocol focuses on mRNA knockdown in zebrafish embryos, it can also be applied to other vertebrates. For complete details on the use and execution of this protocol, please refer to Kushawah et al. (2020). |
format | Online Article Text |
id | pubmed-8715325 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-87153252022-01-06 Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos Hernandez-Huertas, Luis Kushawah, Gopal Diaz-Moscoso, Alejandro Tomas-Gallardo, Laura Moreno-Sanchez, Ismael da Silva Pescador, Gabriel Bazzini, Ariel A. Moreno-Mateos, Miguel A. STAR Protoc Protocol CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo optimized CRISPR-RfxCas13d (CasRx) system. Although the described protocol focuses on mRNA knockdown in zebrafish embryos, it can also be applied to other vertebrates. For complete details on the use and execution of this protocol, please refer to Kushawah et al. (2020). Elsevier 2021-12-23 /pmc/articles/PMC8715325/ /pubmed/35005640 http://dx.doi.org/10.1016/j.xpro.2021.101058 Text en © 2021 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 Hernandez-Huertas, Luis Kushawah, Gopal Diaz-Moscoso, Alejandro Tomas-Gallardo, Laura Moreno-Sanchez, Ismael da Silva Pescador, Gabriel Bazzini, Ariel A. Moreno-Mateos, Miguel A. Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title | Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title_full | Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title_fullStr | Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title_full_unstemmed | Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title_short | Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos |
title_sort | optimized crispr-rfxcas13d system for rna targeting in zebrafish embryos |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715325/ https://www.ncbi.nlm.nih.gov/pubmed/35005640 http://dx.doi.org/10.1016/j.xpro.2021.101058 |
work_keys_str_mv | AT hernandezhuertasluis optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT kushawahgopal optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT diazmoscosoalejandro optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT tomasgallardolaura optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT morenosanchezismael optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT dasilvapescadorgabriel optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT bazziniariela optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos AT morenomateosmiguela optimizedcrisprrfxcas13dsystemforrnatargetinginzebrafishembryos |