Application Of The CRISPR-Cas9 System to Drug Discovery in Breast Cancer

Abstract

To elucidate the functioning of the CRISPR-Cas9 system in the realm of drug discovery, a series of experiments were undertaken. Firstly, CRISPR-Cas9 was employed to facilitate both positive and negative selection screening within human cells. Subsequently, an exploration of genes vital for pluripotent stem cell and cancer cell viability was conducted. Following this, in a melanoma model, a screen was performed to identify genes whose loss contributes to resistance against vemurafenib, a therapeutic agent targeting the mutant protein kinase BRAF. Notably, the top-ranking candidate genes included previously verified genes such as NF1 and MED12, alongside novel discoveries in the form of CUL3, NF2, TADA1, and TADA2B. This investigation revealed a remarkable consistency among distinct guide RNAs directed at the same gene, with an elevated rate of successful hit confirmation. These findings underscore the potential of genome-scale screening employing Cas9, thus substantiating CRISPR-Cas9 as a potent methodology for large-scale sequencing and the selection of target sites.