Combinatorial CRISPR screening is useful for investigating synthetic lethality (SL) gene pairs. Here, we detail the steps for dual-gRNA library construction, with the introduction of two backbones, LentiGuide_DKO and LentiCRISPR_DKO. We describe steps for in vitro screening with 22Rv1-Cas9 and SaOS2-Cas9 cells followed by sequencing and data analysis. By introducing two backbones, we optimized the library construction process, facilitated standard pair-end sequencing, and provided options of screening on cells with or without modification of Cas9 expression.
|State||Published - Sep 16 2022|
Bibliographical noteFunding Information:
The work related to cell culture is done by Dr. Liu’s lab and was supported by NIH R01 CA157429 (X. Liu), R01 CA196634 (X. Liu), R01 CA264652 (X. Liu), R01 CA256893 (X. Liu).
- Biotechnology and bioengineering
- High Throughput Screening
- Molecular Biology
ASJC Scopus subject areas
- Neuroscience (all)
- Biochemistry, Genetics and Molecular Biology (all)
- Immunology and Microbiology (all)