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Integrated Target Identification and Validation Platform

Overview

 

 

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