GDesigner: computational design of synthetic gRNAs for Cas12a-based transcriptional repression in mammalian cells.

ABSTRACT: Synthetic networks require complex intertwined genetic regulation often relying on transcriptional activation or repression of target genes. CRISPRi-based transcription factors facilitate the programmable modulation of endogenous or synthetic promoter activity and the process can be optimised by using software to select appropriate gRNAs and limit non-specific gene modulation. Here, we develop a computational software pipeline, gDesigner, that enables the automated selection of orthogonal gRNAs with minimized off-target effects and promoter crosstalk. We next engineered a Lachnospiraceae bacterium Cas12a (dLbCas12a)-based repression system that downregulates target gene expression by means of steric hindrance of the cognate promoter. Finally, we generated a library of orthogonal synthetic

SUBMITTER: Crone MA 

PROVIDER: S-EPMC9481559 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature