1 Generation of cell lines with inducible gene overexpression or knockdown via lentiviral system
2 Generation of cell lines with gene knockout or knockin via CRISPR-Cas9 system
Knockout via transient transfection, followed by 2D colony formation assay and drug response profiling
Knockout via lentiviral CRISPR/Cas9 system
Introduction of gene mutation or endogenous tagging by CRISPR/Cas9 knockin
3 In vitro CRISPR screen
Overview of in vitro CRISPR screen workflow
In-house pipeline for customized design
Cancer cell line based positive selection or negative selection screen to identify mechanism of drug resistance or essential genes
CRISPR screen in cancer cell and T cell co-culture to identify regulators of cancer cell response to T cell cytotoxicity
Bioinformatics workflow for screening data
4 In vivo CRISPR screen
Overview of in vivo CRISPR screen workflow
In vivo CRISPR screen for cancer immunotherapy modulators in syngeneic tumor models
Case study: in vivo CRISPR screen with a library derived from tumor evolution under immune selection identified a gene that is required for PD-1 independent T cell cytotoxicity
