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CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
Cpf1 represents a novel single RNA‐guided CRISPR/Cas endonuclease system suitable for genome editing with distinct features compared with Cas9. We demonstrate the functionality of three Cpf1 orthologues – Acidaminococcus spp. BV3L6 (AsCpf1), Lachnospiraceae bacterium ND2006 (LbCpf1) and Francisella...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5836994/ https://www.ncbi.nlm.nih.gov/pubmed/28886218 http://dx.doi.org/10.1002/yea.3278 |
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author | Verwaal, René Buiting‐Wiessenhaan, Nathalie Dalhuijsen, Sacha Roubos, Johannes A. |
author_facet | Verwaal, René Buiting‐Wiessenhaan, Nathalie Dalhuijsen, Sacha Roubos, Johannes A. |
author_sort | Verwaal, René |
collection | PubMed |
description | Cpf1 represents a novel single RNA‐guided CRISPR/Cas endonuclease system suitable for genome editing with distinct features compared with Cas9. We demonstrate the functionality of three Cpf1 orthologues – Acidaminococcus spp. BV3L6 (AsCpf1), Lachnospiraceae bacterium ND2006 (LbCpf1) and Francisella novicida U112 (FnCpf1) – for genome editing of Saccharomyces cerevisiae. These Cpf1‐based systems enable fast and reliable introduction of donor DNA on the genome using a two‐plasmid‐based editing approach together with linear donor DNA. LbCpf1 and FnCpf1 displayed editing efficiencies comparable with the CRISPR/Cas9 system, whereas AsCpf1 editing efficiency was lower. Further characterization showed that AsCpf1 and LbCpf1 displayed a preference for their cognate crRNA, while FnCpf1‐mediated editing with similar efficiencies was observed using non‐cognate crRNAs of AsCpf1 and LbCpf1. In addition, multiplex genome editing using a single LbCpf1 crRNA array is shown to be functional in yeast. This work demonstrates that Cpf1 broadens the genome editing toolbox available for Saccharomyces cerevisiae. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd. |
format | Online Article Text |
id | pubmed-5836994 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-58369942018-03-12 CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae Verwaal, René Buiting‐Wiessenhaan, Nathalie Dalhuijsen, Sacha Roubos, Johannes A. Yeast Research Articles Cpf1 represents a novel single RNA‐guided CRISPR/Cas endonuclease system suitable for genome editing with distinct features compared with Cas9. We demonstrate the functionality of three Cpf1 orthologues – Acidaminococcus spp. BV3L6 (AsCpf1), Lachnospiraceae bacterium ND2006 (LbCpf1) and Francisella novicida U112 (FnCpf1) – for genome editing of Saccharomyces cerevisiae. These Cpf1‐based systems enable fast and reliable introduction of donor DNA on the genome using a two‐plasmid‐based editing approach together with linear donor DNA. LbCpf1 and FnCpf1 displayed editing efficiencies comparable with the CRISPR/Cas9 system, whereas AsCpf1 editing efficiency was lower. Further characterization showed that AsCpf1 and LbCpf1 displayed a preference for their cognate crRNA, while FnCpf1‐mediated editing with similar efficiencies was observed using non‐cognate crRNAs of AsCpf1 and LbCpf1. In addition, multiplex genome editing using a single LbCpf1 crRNA array is shown to be functional in yeast. This work demonstrates that Cpf1 broadens the genome editing toolbox available for Saccharomyces cerevisiae. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd. John Wiley and Sons Inc. 2017-11-12 2018-02 /pmc/articles/PMC5836994/ /pubmed/28886218 http://dx.doi.org/10.1002/yea.3278 Text en © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd. This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial‐NoDerivs (http://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Research Articles Verwaal, René Buiting‐Wiessenhaan, Nathalie Dalhuijsen, Sacha Roubos, Johannes A. CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae |
title | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
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title_full | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
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title_fullStr | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
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title_full_unstemmed | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
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title_short | CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae
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title_sort | crispr/cpf1 enables fast and simple genome editing of saccharomyces cerevisiae |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5836994/ https://www.ncbi.nlm.nih.gov/pubmed/28886218 http://dx.doi.org/10.1002/yea.3278 |
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