Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes
Journal article, 2017

Bioinformatics tools to design guide-RNAs (gRNAs) in Clustered Regularly Interspaced Short Palindromic Repeats systems mostly focused on minimizing off-targeting to enhance efficacy of genome editing. However, there are circumstances in which off-targeting might be desirable to target multiple genes simultaneously with a single gRNA. We termed these gRNAs as promiscuous gRNAs. Here, we present a computational workflow to identify promiscuous gRNAs that putatively bind to the region of interest for a defined list of genes in a genome. We experimentally validated two promiscuous gRNA for gene deletion, one targeting FAA1 and FAA4 and one targeting PLB1 and PLB2, thus demonstrating that multiplexed genome editing through design of promiscuous gRNA can be performed in a time and cost-effective manner.

computational tool

synthetic biology

CRISPR

Author

Raphael Ferreira

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk Foundation

Francesco Gatto

University of California at Los Angeles

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jens B Nielsen

Novo Nordisk Foundation

Technical University of Denmark (DTU)

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

FEBS Letters

0014-5793 (ISSN) 18733468 (eISSN)

Vol. 591 20 3288-3295

Subject Categories

Biochemistry and Molecular Biology

Infrastructure

Chalmers Infrastructure for Mass spectrometry

DOI

10.1002/1873-3468.12835

PubMed

28884816

More information

Latest update

5/26/2023