Home > Browse Issues > Vol.40 No.9
Research of CRISPR/Cas9 Technique to Knock Out Bloom Helicase Gene in Human Prostate Cancer PC-3 Cells
Chen Kun1,2,3, Xu Houqiang1*, Zhao Jiafu1, Duan Zhiqiang1, Wu Ping1
1Key Laboratory of Animal Genetics, Breeding and Production in the Pleteau Mountains Region, Ministry of Education, Guizhou University, Guiyang 550025, China; 2College of Life Science, Guizhou University, Guiyang 550025, China; 3Guizhou Province People’s Hospital, Guiyang 550003, China
Abstract: BLM helicase is an important member of the human RecQ helicase family， which plays a crucial role in DNA replication， repair， recombinant， transcription， telomere maintenance and other cell metabolism. The BLM helicase gene is associated with susceptibility to prostate cancer and it is also a candidate gene for treatment of prostate cancer. In this study， the five Cas9/SgRNA vectors were designed， and we selected the most active SgRNA3 vector by using T7E1 enzyme. Two vectors of Cas9/SgRNA3 and donor vector were jointly transduced into human prostate cancer PC-3 cell by LTX liposomes. BLa and Puro resistance screenings and fluorescent protein markers were used to identify cells with positive markers. The BLM helicase gene in PC-3 cells of prostate cancer was successfully knocked out detected by Western blot and RTq-PCR tests. This study can provide the important scientific data which play a role of the BLM helicase in prostate cancer.
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