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.
Candidate Gene Screening Principle
The deletion of gene itself has no significant effect on cell proliferation, but has a significant inhibitory effect on cell proliferation when combined with drugs, which may be used as a potential target for sensitization of chemotherapeutic drugs.
According to the candidate sensitization genes, combined with the disease key gene database and the high connotation functional gene screening platform (HCS), we screened the genes which had not been reported in the studied tumors, and could significantly increase the sensitivity of cells to drugs, and completed the effects of the gene on cell proliferation, apoptosis, cycle, clone formation and so on, and elucidated the molecular mechanism of the function of the gene.
Mature and reliable tumor genome-wide screening platform: rich project experience, participate in major projects, and publish high-level cooperation results.
The analysis contents and methods based on high-quality data analysis, pre-research and verification are indeed mature and feasible.
Creative Proteomics leads a rich team that uses the most rigorous experimental solutions and internationally recognized experimental techniques for Genome-wide CAS9 Screening to obtain your customized information. Use the most accurate experimental results to help you better carry out tumor-related scientific research.
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