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.

computational tool

synthetic biology



Raphael Ferreira

Chalmers, Biologi och bioteknik, Systembiologi

Novo Nordisk Fonden

Francesco Gatto

University of California at Los Angeles

Chalmers, Biologi och bioteknik, Systembiologi

Jens B Nielsen

Novo Nordisk Fonden

Danmarks Tekniske Universitet (DTU)

Chalmers, Biologi och bioteknik, Systembiologi

FEBS Letters

0014-5793 (ISSN) 18733468 (eISSN)

Vol. 591 20 3288-3295


Biokemi och molekylärbiologi


Chalmers infrastruktur för masspektrometri





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