Feasibility of CRISPR-Cas9-Based In Vitro Drug Target Identification for Personalized Prostate Cancer Medicine
Technical Report,01 Sep 2016,31 Aug 2017
University of Tampere Tampere Finland
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This study tests the feasibility of using CRISPR-Cas9 to introduce patient-derived clonal truncal mutations into prostate cell lines in order to study their potential druggability. We aimed to establish a positive control cell line by introducing a known PIK3CA mutation that is druggable using available PI3K pathway inhibitors. However, using immortalized normal prostate epithelial cell line RWPE-1 we had significant problems getting CRISPR working efficiently. We suspected this being due to the chromatin status of PIK3CA locus in these cells, as the gene in very lowly expressed. Thus, we have moved on to creating the actual patient-derived clonal truncal mutations in this cell line. Currently we are in the process of creating clonal cell lines and hope to be able to sequence-verify targeted mutagenesis and then proceed to drug sensitivity testing.
- Medicine and Medical Research