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

René Verwaal, Nathalie Buiting-Wiessenhaan, Sacha Dalhuijsen, Johannes A. Roubos

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)201-211
Number of pages11
JournalYeast
Volume35
Issue number2
DOIs
Publication statusPublished - Feb 1 2018

Keywords

  • CRISPR/Cas9
  • CRISPR/Cpf1
  • Saccharomyces cerevisiae
  • genome editing
  • single Cpf1 crRNA array

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics

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