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Analysis of a Cas12a-based gene-drive system in budding yeast
The discovery and adaptation of CRISPR/Cas systems within molecular biology has provided advances across biological research, agriculture and human health. Genomic manipulation through use of a CRISPR nuclease and programmed guide RNAs has become a common and widely accessible practice. The identifi...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Microbiology Society
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749140/ https://www.ncbi.nlm.nih.gov/pubmed/35024561 http://dx.doi.org/10.1099/acmi.0.000301 |
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author | Lewis, Isabel C. Yan, Yao Finnigan, Gregory C. |
author_facet | Lewis, Isabel C. Yan, Yao Finnigan, Gregory C. |
author_sort | Lewis, Isabel C. |
collection | PubMed |
description | The discovery and adaptation of CRISPR/Cas systems within molecular biology has provided advances across biological research, agriculture and human health. Genomic manipulation through use of a CRISPR nuclease and programmed guide RNAs has become a common and widely accessible practice. The identification and introduction of new engineered variants and orthologues of Cas9 as well as alternative CRISPR systems such as the type V group have provided additional molecular options for editing. These include distinct PAM requirements, staggered DNA double-strand break formation, and the ability to multiplex guide RNAs from a single expression construct. Use of CRISPR/Cas has allowed for the construction and testing of a powerful genetic architecture known as a gene drive within eukaryotic model systems. Our previous work developed a drive within budding yeast using Streptococcus pyogenes Cas9. Here, we installed the type V Francisella novicida Cas12a (Cpf1) nuclease gene and its corresponding guide RNA to power a highly efficient artificial gene drive in diploid yeast. We examined the consequence of altering guide length or introduction of individual mutational substitutions to the crRNA sequence. Cas12a-dependent gene-drive function required a guide RNA of at least 18 bp and could not tolerate most changes within the 5′ end of the crRNA. |
format | Online Article Text |
id | pubmed-8749140 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Microbiology Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-87491402022-01-11 Analysis of a Cas12a-based gene-drive system in budding yeast Lewis, Isabel C. Yan, Yao Finnigan, Gregory C. Access Microbiol Research Articles The discovery and adaptation of CRISPR/Cas systems within molecular biology has provided advances across biological research, agriculture and human health. Genomic manipulation through use of a CRISPR nuclease and programmed guide RNAs has become a common and widely accessible practice. The identification and introduction of new engineered variants and orthologues of Cas9 as well as alternative CRISPR systems such as the type V group have provided additional molecular options for editing. These include distinct PAM requirements, staggered DNA double-strand break formation, and the ability to multiplex guide RNAs from a single expression construct. Use of CRISPR/Cas has allowed for the construction and testing of a powerful genetic architecture known as a gene drive within eukaryotic model systems. Our previous work developed a drive within budding yeast using Streptococcus pyogenes Cas9. Here, we installed the type V Francisella novicida Cas12a (Cpf1) nuclease gene and its corresponding guide RNA to power a highly efficient artificial gene drive in diploid yeast. We examined the consequence of altering guide length or introduction of individual mutational substitutions to the crRNA sequence. Cas12a-dependent gene-drive function required a guide RNA of at least 18 bp and could not tolerate most changes within the 5′ end of the crRNA. Microbiology Society 2021-12-17 /pmc/articles/PMC8749140/ /pubmed/35024561 http://dx.doi.org/10.1099/acmi.0.000301 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License. |
spellingShingle | Research Articles Lewis, Isabel C. Yan, Yao Finnigan, Gregory C. Analysis of a Cas12a-based gene-drive system in budding yeast |
title | Analysis of a Cas12a-based gene-drive system in budding yeast |
title_full | Analysis of a Cas12a-based gene-drive system in budding yeast |
title_fullStr | Analysis of a Cas12a-based gene-drive system in budding yeast |
title_full_unstemmed | Analysis of a Cas12a-based gene-drive system in budding yeast |
title_short | Analysis of a Cas12a-based gene-drive system in budding yeast |
title_sort | analysis of a cas12a-based gene-drive system in budding yeast |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8749140/ https://www.ncbi.nlm.nih.gov/pubmed/35024561 http://dx.doi.org/10.1099/acmi.0.000301 |
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