CRISPR/Cas9 system

The sgRNA-mediated DNA cleavage mechanism is a defense mechanism found in bacteria. It can edit Target genes by designing the sgRNA. As an emerging method of DNA manipulation, the technology has prompted gene editing selected MIT Technology Review 2014 annual top ten technological breakthroughs. CRISPR / Cas9 gene editing system is easy operating, wide usable, high efficiency, and can knock out multiple genes.

CRISPR SCREEN

From 2014, The large-scale gene knockout screening, which combines the CRISPR / Cas9 system with next-generation sequencing, has greatly promoted the selection of molecular markers related to essential genes, drug responses and drug resistance in specific cell types and treatment conditions. The CRISPR screening assay is characterized by high throughput, high efficiency, and low cost compared to conventional low-throughput screening, early small RNA knock-down screening (RNAi), and mutation profiling. These advantages make CRISPR screening with unparalleled potential in the field of precision medicine.

In-vitro CRISPR screen

GV20 Oncotherapy has already built a highly parallel platform for In-vitro CRISPR screen platform. Advances in pooled screening have made it possible to readily evaluate mammalian gene phenotypic function at genome-scale. Our In-vitro CRISPR screen includes cell growth CRISPR screen, drug treatment CRISPR screen and cancer cells and immune cell co-culture.
Tumor cell growth screen provide a valuable addition to the methods available for novel drug target identification and validation. Drug treatment screen can be used to investigate the mechanism of drug resistance and drug combinations for cancer therapy. tumor and immune cell co-culture assays can be used to identify the genes essential in tumors for the effector function of immune cells.

In-vivo CRISPR Screen

GV20 Oncotherapy simultaneously builds In-vivo CRISPR screen platform, including construction of Syngeneic Tumor Mouse Mode, transplanted into immunocompromised mice, genetic screen, and downstream data analysis. GV20 Oncotherapy has successfully performed in-vivo CRISPR screen with the treatment of PDL-1 inhibitor.

MAGeCK-VISPR >10000 downloads

Li, Xu et al, Genome Biol 2014 Li, Koster et al, Genome Biol 2015
CRISPR SCREEN data quality control, analysis, and visualization

Algorithm / Tools
sgRNA design optimization

Xu et al., Genome Biol. 2015
sgRNA design optimization based on target sequence content

NEST

Jiang et al, Genome Biol 2015
CRISPR SCREEN result optimization with integrated data of protein-protein interaction

CRISPR-DO

Ma et al., CRISPR-DO for genome-wide CRISPR design and optimization. Bioinformatics. 2016
Whole genome CRISPR sgRNA design in several species