Journal
FEBS LETTERS
Volume 591, Issue 20, Pages 3288-3295Publisher
WILEY
DOI: 10.1002/1873-3468.12835
Keywords
computational tool; CRISPR; synthetic biology
Funding
- Novo Nordisk Foundation
- Knut and Alice Wallenberg Foundation
- Swedish Foundation for Strategic Research
- NNF Center for Biosustainability [Yeast Cell Factories] Funding Source: researchfish
- Novo Nordisk Fonden [NNF10CC1016517] Funding Source: researchfish
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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.
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