The Creative Proteomics team used the CAS9 whole-genome knockout library, combined with high-throughput sequencing and bioinformatics, to analyze and screen key gene sets related to tolerance sensitivity under different processing factors, such as anoxic stress, antineoplastic drugs, etc. Functional verification and mechanism analysis of the selected key sensitization genes were carried out to provide preliminary data for the determination of sensitization targets of antineoplastic drugs.

Creative Proteomics's CRISPR/Cas9-sgRNA (single guide RNA) whole-genome library CRISPR-PoolTMKOUT is a genomic-wide CRISPR knockout library that can knock out any coding gene and part of miRNA in the human genome. The library contains 6 sgRNA for each coding gene, 4 sgRNA, for each miRNA and 1000 non-targeted control sgRNA.

In order to screen the sensitizing genes that can increase the sensitivity of cells to the target drug (or target treatment factors), the cells were infected with lentivirus with CAS9 whole genome knockout library and screened by puromycin. After the successful infection of the library, the target drugs were added or treated under other conditions, and the cells were collected at different time points. The genomic DNA, was extracted and amplified by universal primers designed by sgRNA. The sgRNA components were detected by high-throughput sequencing, and the candidate sensitizing genes were obtained according to the sgRNA enrichment of cell samples at different time points.

Genome-wide Screening of New Tumor Drug Targets

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