FnCpf1: a novel and efficient genome editing tool for Saccharomyces cerevisiae

Michal A. Swiat, Sofia Dashko, Maxime den Ridder, Melanie Wijsman, John van der Oost, Jean Marc Daran, Pascale Daran-Lapujade*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

39 Citations (Scopus)

Abstract

Cpf1 is a new class II family of CRISPR-Cas RNA-programmable endonucleases with unique features that make it a very attractive alternative or complement to Cas9 for genome engineering. Using constitutively expressed Cpf1 from Francisella novicida, the present study demonstrates that FnCpf1 can mediate RNA-guided DNA cleavage at targeted genomic loci in the popular model and industrial yeast Saccharomyces cerevisiae. FnCpf1 very efficiently and precisely promoted repair DNA recombination with efficiencies up to 100%. Furthermore, FnCpf1 was shown to introduce point mutations with high fidelity. While editing multiple loci with Cas9 is hampered by the need for multiple or complex expression constructs, processing itself a customized CRISPR array FnCpf1 was able to edit four genes simultaneously in yeast with a 100% efficiency. A remarkable observation was the unexpected, strong preference of FnCpf1 to cleave DNA at target sites harbouring 5'-TTTV-3' PAM sequences, a motif reported to be favoured by Cpf1 homologs of Acidaminococcus and Lachnospiraceae. The present study supplies several experimentally tested guidelines for crRNA design, as well as plasmids for FnCpf1 expression and easy construction of crRNA expression cassettes in S. cerevisiae. FnCpf1 proves to be a powerful addition to S. cerevisiae CRISPR toolbox.
Original languageEnglish
Pages (from-to)12585-12598
JournalNucleic acids research
Volume45
Issue number21
DOIs
Publication statusPublished - 1 Dec 2017

Fingerprint Dive into the research topics of 'FnCpf1: a novel and efficient genome editing tool for Saccharomyces cerevisiae'. Together they form a unique fingerprint.

  • Cite this

    Swiat, M. A., Dashko, S., den Ridder, M., Wijsman, M., van der Oost, J., Daran, J. M., & Daran-Lapujade, P. (2017). FnCpf1: a novel and efficient genome editing tool for Saccharomyces cerevisiae. Nucleic acids research, 45(21), 12585-12598. https://doi.org/10.1093/nar/gkx1007