CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces cerevisiae

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...

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Bibliographic Details
Published in:Yeast (Chichester, England) Vol. 35; no. 2; pp. 201 - 211
Main Authors: Verwaal, René, Buiting‐Wiessenhaan, Nathalie, Dalhuijsen, Sacha, Roubos, Johannes A.
Format: Journal Article
Language:English
Published: England Wiley Subscription Services, Inc 01-02-2018
John Wiley and Sons Inc
Subjects:
CRISPR
CRISPR-Cas Systems
CRISPR/Cas9
CRISPR/Cpf1
Deoxyribonucleic acid
DNA
DNA, Fungal - genetics
Endonuclease
Fungal Proteins - genetics
Fungal Proteins - metabolism
Gene Editing
Gene Expression Regulation, Fungal
Genome editing
Genomes
Ribonucleic acid
RNA
RNA, Fungal
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
single Cpf1 crRNA array
Yeast
CRISPR/Cpf1
single Cpf1 crRNA array
genome editing
CRISPR/Cas9
Saccharomyces cerevisiae
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Summary: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.
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ISSN:0749-503X
1097-0061
DOI:10.1002/yea.3278