CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish
Summary: Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166720301957 |
| _version_ | 1818921365432958976 |
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| author | João Pedro Amorim Renata Bordeira-Carriço Ana Gali-Macedo Chiara Perrod José Bessa |
| author_facet | João Pedro Amorim Renata Bordeira-Carriço Ana Gali-Macedo Chiara Perrod José Bessa |
| author_sort | João Pedro Amorim |
| collection | DOAJ |
| description | Summary: Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and Cas9 mRNA, followed by microinjection in zebrafish embryos and subsequent genotype screening for the establishment of a mutant line. The versatility and efficiency of this pipeline makes the generation of zebrafish models a widely used approach in functional genetics.For complete details on the use and execution of this protocol, please refer to Amorim et al. (2020). |
| first_indexed | 2024-12-20T01:36:29Z |
| format | Article |
| id | doaj.art-b14375d777fd483b997e27ff75d9360c |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-20T01:36:29Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-b14375d777fd483b997e27ff75d9360c2022-12-21T19:58:00ZengElsevierSTAR Protocols2666-16672020-12-0113100208CRISPR-Cas9-Mediated Genomic Deletions Protocol in ZebrafishJoão Pedro Amorim0Renata Bordeira-Carriço1Ana Gali-Macedo2Chiara Perrod3José Bessa4i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugali3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugali3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugali3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal; Corresponding authori3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; IBMC – Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal; Corresponding authorSummary: Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and Cas9 mRNA, followed by microinjection in zebrafish embryos and subsequent genotype screening for the establishment of a mutant line. The versatility and efficiency of this pipeline makes the generation of zebrafish models a widely used approach in functional genetics.For complete details on the use and execution of this protocol, please refer to Amorim et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166720301957CRISPRGeneticsGenomicsModel Organisms |
| spellingShingle | João Pedro Amorim Renata Bordeira-Carriço Ana Gali-Macedo Chiara Perrod José Bessa CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish STAR Protocols CRISPR Genetics Genomics Model Organisms |
| title | CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish |
| title_full | CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish |
| title_fullStr | CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish |
| title_full_unstemmed | CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish |
| title_short | CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish |
| title_sort | crispr cas9 mediated genomic deletions protocol in zebrafish |
| topic | CRISPR Genetics Genomics Model Organisms |
| url | http://www.sciencedirect.com/science/article/pii/S2666166720301957 |
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