Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes
Artikel i vetenskaplig tidskrift, 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.

CRISPR

synthetic biology

computational tool

Författare

Raphael Ferreira

Chalmers, Biologi och bioteknik, Systembiologi

Francesco Gatto

Chalmers, Biologi och bioteknik, Systembiologi

Jens B Nielsen

Chalmers, Biologi och bioteknik, Systembiologi

FEBS Letters

0014-5793 (ISSN)

Vol. 591 20 3288-3295

Ämneskategorier

Biokemi och molekylärbiologi

Infrastruktur

Chalmers infrastruktur för masspektrometri

DOI

10.1002/1873-3468.12835

PubMed

28884816

Mer information

Skapat

2017-11-17