Klk gene editing using crispr technology in prostate cancer cell lines for cancer therapy

Prostate cancer (PC) has been accounted for as the second leading cause of death by cancer in men. Internal and external factors, such as genetic mutation, ethnicity, age, diet, etc., are observed to have been the causative elements for cancer to occur. Serine proteases, that are encoded by Kallikrein (KLK) gene family, show a remarkable impact on PC and some are introduced as PC biomarkers, such as KLK3 and KLK2, as both androgen-dependent are detected in the bloodstream when PC occurs. However, due to androgen dependence and low specificity, they are not quite reliable. In this study, we focused on KLK5 gene mutation in different PC androgen-independent cell lines, PC3, and DU145 cell lines to observe the effect of mutation of PC tumorigenesis. To monitor the effect of the mutation, the KLK5 gene's detected mutation point was knocked out using a genome-editing tool. Gene-editing technology has opened a new path to the scientists' investigation for designing the method that can benefit cancer treatment with the lowest side effects in the long term. In this study, CRISPR/Cas9 gene-editing technology was used to knock out c.245C>A (p.A82D) mutation in the KLK gene which was detected and confirmed by Sanger sequencing. To evaluate the performed gene editing, various cytotoxicity assays, such as cell viability with trypan blue, MTT assay, and colony formation was carried using Doxorubicin and PARP inhibitor which showed a promising result of CRISPR system as a future prostate cancer treatment.