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

Autor: Verwaal R; DSM Biotechnology Center, Alexander Fleminglaan 1, 2600, MA, Delft, the Netherlands., Buiting-Wiessenhaan N; DSM Biotechnology Center, Alexander Fleminglaan 1, 2600, MA, Delft, the Netherlands., Dalhuijsen S; DSM Biotechnology Center, Alexander Fleminglaan 1, 2600, MA, Delft, the Netherlands., Roubos JA; DSM Biotechnology Center, Alexander Fleminglaan 1, 2600, MA, Delft, the Netherlands.
Jazyk: angličtina
Zdroj: Yeast (Chichester, England) [Yeast] 2018 Feb; Vol. 35 (2), pp. 201-211. Date of Electronic Publication: 2017 Nov 12.
DOI: 10.1002/yea.3278
Abstrakt: 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.
(© 2017 The Authors. Yeast published by John Wiley & Sons, Ltd.)
Databáze: MEDLINE
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