Creating a dual-guide RNA library to test pairs of genes that may cause cell death when turned off together

Combinatorial CRISPR screening can be utilized to investigate synthetic lethality gene pairs using two newly developed backbones.

• The introduction of LentiGuide_DKO and LentiCRISPR_DKO backbones optimized the dual-gRNA library construction process.
• Screening was performed on 22Rv1-Cas9 and SaOS2-Cas9 cells.
• Standard pair-end sequencing was facilitated through the new backbones.
• Options for screening were provided for cells with or without modified Cas9 expression.
• The steps for data analysis following sequencing were outlined.

Simplified