겨울 심포지움
2018겨울초록
발표자 및 발표 내용
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공동저자
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As an important key to cancer treatment, signlaing pathways regulated by kinasses are the targets of most cancer drugs. However, targeted anticancer agents are facing limitations due to their resistance and low popularity and efficiencies. The kinase genes were edited using CRISPR-Cas9 technology and target deconvolution was performed by screening from various omics perspectives to find an aberrantly activated signaling pathway and an unintended target of kinase inhibitor.
Four kinases (ERK2, PIK3CA PLK1, and PAK4) of HCT116 cells, were knocked out using CRISPR-Cas9 to obtain cell lines. Proteins were extracted from these knockout cell lines and tryptic digestion followed by labeling using TMT reagent. We performed global profiling and phosphopeptides enrichment using IMAC method for the labeled proteins. As a result, a total of 7,500 proteins, 4200 phosphoproteins and 10877 phosphosites were identified by LC-MS / MS.
Results of GOBP enriched for DEPs (Differentially Expressed Proteins) and DPPs (Differentially Expressed Phosphoproteins) in each knockout cell line were obtained and a model of protein-protein interaction network was established.
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