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A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh
The introduction of CRISPR-Cas9 technology for targeted mutagenesis has revolutionized reverse genetics and made genome editing a realistic option in many model organisms. One of the difficulties with this technique is screening for mutations in large numbers of samples. Many screening approaches fo...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9580339/ https://www.ncbi.nlm.nih.gov/pubmed/29939088 http://dx.doi.org/10.2144/btn-2018-0012 |
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author | VanLeuven, Ariel J Park, Sungdae Menke, Douglas B Lauderdale, James D |
author_facet | VanLeuven, Ariel J Park, Sungdae Menke, Douglas B Lauderdale, James D |
author_sort | VanLeuven, Ariel J |
collection | PubMed |
description | The introduction of CRISPR-Cas9 technology for targeted mutagenesis has revolutionized reverse genetics and made genome editing a realistic option in many model organisms. One of the difficulties with this technique is screening for mutations in large numbers of samples. Many screening approaches for identifying CRISPR-Cas9 mutants have been published; however, in practice these methods are time consuming, expensive, or often yield false positives. This report describes a PCR-based screening approach using non-denaturing PAGE. This approach does not depend on the formation of heteroduplexes and reliably detects changes as small as 1 base-pair (bp) in nucleic acid length at the target site. This approach can be used to identify novel mutations and is also useful as a routine genotyping method. |
format | Online Article Text |
id | pubmed-9580339 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
record_format | MEDLINE/PubMed |
spelling | pubmed-95803392022-10-19 A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh VanLeuven, Ariel J Park, Sungdae Menke, Douglas B Lauderdale, James D Biotechniques Article The introduction of CRISPR-Cas9 technology for targeted mutagenesis has revolutionized reverse genetics and made genome editing a realistic option in many model organisms. One of the difficulties with this technique is screening for mutations in large numbers of samples. Many screening approaches for identifying CRISPR-Cas9 mutants have been published; however, in practice these methods are time consuming, expensive, or often yield false positives. This report describes a PCR-based screening approach using non-denaturing PAGE. This approach does not depend on the formation of heteroduplexes and reliably detects changes as small as 1 base-pair (bp) in nucleic acid length at the target site. This approach can be used to identify novel mutations and is also useful as a routine genotyping method. 2018-06 /pmc/articles/PMC9580339/ /pubmed/29939088 http://dx.doi.org/10.2144/btn-2018-0012 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under the Creative Commons Attribution 4.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) |
spellingShingle | Article VanLeuven, Ariel J Park, Sungdae Menke, Douglas B Lauderdale, James D A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title | A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title_full | A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title_fullStr | A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title_full_unstemmed | A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title_short | A PAGE screening approach for identifying CRISPR-Cas9-induced mutations in zebrafi sh |
title_sort | page screening approach for identifying crispr-cas9-induced mutations in zebrafi sh |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9580339/ https://www.ncbi.nlm.nih.gov/pubmed/29939088 http://dx.doi.org/10.2144/btn-2018-0012 |
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