Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes

Raphael Ferreira, Francesco Gatto, Jens Nielsen

Research output: Contribution to journalJournal articleResearchpeer-review

3 Downloads (Pure)

Abstract

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.
Original languageEnglish
JournalFEBS Letters
Volume591
Issue number20
Pages (from-to)3288-3295
ISSN0014-5793
DOIs
Publication statusPublished - 2017

Keywords

  • CRISPR
  • Computational tool
  • Synthetic biology

Fingerprint

Dive into the research topics of 'Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes'. Together they form a unique fingerprint.

Cite this