Journal article
Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes
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.
Language: | English |
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Publisher: | Wiley |
Year: | 2017 |
Pages: | 3288-3295 |
ISSN: | 18733468 and 00145793 |
Types: | Journal article |
DOI: | 10.1002/1873-3468.12835 |
ORCIDs: | 0000-0001-9881-6232 |
Acyl Coenzyme A Base Sequence CRISPR-Cas Systems Clustered Regularly Interspaced Short Palindromic Repeats Coenzyme A Ligases Computational Biology FAA4 protein, S cerevisiae Faa1 protein, S cerevisiae Gene Deletion Gene Editing Gene Expression Genome, Fungal Lysophospholipase Membrane Proteins PLB1 protein, S cerevisiae PLB2 protein, S cerevisiae RNA, Guide, CRISPR-Cas Systems Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins computational tool synthetic biology