CRISPRi-FGP: a web-based genome-scale CRISPRi sgRNA design tool for prokaryotes

Abstract

ABSTRACTCRISPRi screening is emerging as a powerful strategy for functional genomic study. However, off-target effects that result from mismatch tolerance between sgRNAs and targets are regarded as the major problem in CRISPRi applications. To avoid off-target binding, here we propose a web-based tool named CRISPRi-Functional Genomics for Prokaryotes (CRISPRi-FGP) for genome-scale sgRNA library design for prokaryotic organisms (http://www.thu-big.net/sgRNA_design/). For this tool, we established a powerful quality control principle that identifies off-target hits with high accuracy. Moreover, we established a built-in database containing 1,300 common prokaryotes as a design source. In addition, CRISPRi-FGP also supports sgRNA design for new organisms using uploaded reference files. Finally, we found that compared to existing sgRNA design tools, CRISPRi-FGP could identify off-target binding hits with higher precision.