Multiplex genome editing of microorganisms using CRISPR-Cas

Belén Adiego-Pérez; Paola Randazzo; Jean Marc Daran; René Verwaal; Johannes A Roubos; Pascale Daran-Lapujade; John van der Oost

doi:10.1093/femsle/fnz086

Multiplex genome editing of microorganisms using CRISPR-Cas

FEMS Microbiol Lett. 2019 Apr 1;366(8):fnz086. doi: 10.1093/femsle/fnz086.

Authors

Belén Adiego-Pérez¹, Paola Randazzo², Jean Marc Daran², René Verwaal³, Johannes A Roubos³, Pascale Daran-Lapujade², John van der Oost¹

Affiliations

¹ Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, 6708 WE Wageningen, The Netherlands.
² Department of Biotechnology, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, The Netherlands.
³ DSM Biotechnology Center, Alexander Fleminglaan 1, 2613 AX Delft, The Netherlands.

Abstract

Microbial production of chemical compounds often requires highly engineered microbial cell factories. During the last years, CRISPR-Cas nucleases have been repurposed as powerful tools for genome editing. Here, we briefly review the most frequently used CRISPR-Cas tools and describe some of their applications. We describe the progress made with respect to CRISPR-based multiplex genome editing of industrial bacteria and eukaryotic microorganisms. We also review the state of the art in terms of gene expression regulation using CRISPRi and CRISPRa. Finally, we summarize the pillars for efficient multiplexed genome editing and present our view on future developments and applications of CRISPR-Cas tools for multiplex genome editing.

Keywords: CRISPR-Cas; Cas12a; Cas9; cell factories; genome editing; multiplex.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Bacteria / genetics*
CRISPR-Cas Systems*
Gene Editing / methods*
Gene Editing / trends
Gene Expression Regulation, Bacterial*
Industrial Microbiology / methods*