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

Abstract Cpf1 represents a novel single RNA‐guided CRISPR/Cas endonuclease system suitable for genome editing with distinct features compared to Cas9. We demonstrate the functionality of three Cpf1 orthologues ‐ Acidaminococcus spp. BV3L6 (AsCpf1), Lachnospiraceae bacterium ND2006 (LbCpf1), 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 to that of the CRISPR/Cas9 system, whereas AsCpf1 editing efficiency was lower. Further characterization showed that AsCpf1 and LbCpf1 displayed 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.
Source: Yeast - Category: Molecular Biology Authors: Tags: Research Article Source Type: research