Knock-out of androgen receptor gene based on CRISPR/Cas9: an experimental study

Objective To investigate the knock-out of androgen receptor (AR) gene at the cellular level, and to provide cell lines for the study of AR pathway in prostate diseases. Methods The sgRNA target sequence for AR was designed by online software. The designed sgRNA target sequence was cloned into PX330 vector and verified by sequencing. The cloned CRISPR-AR vector was transfected into T293 cells, and the DNA was extracted. PCR, restriction enzyme digestion and sequencing were used to identify the gene knockout efficiency. Results Sequencing of CRISPR-AR vector verified that the plasmid vector was successfully constructed. 293T cell transfection and restriction enzyme digestion showed that the CRISPR-AR vector had a knockout efficiency of 95.3%. Conclusion Based on CRISPR/Cas9, the established CRISPR-AR vector can be used to knock out AR genes in cells, which can be further used for the construction of AR gene knockout cell lines. Key words: Receptor, androgen; Benign prostatic hyperplasia; Prostatic neoplasms; CRISPR/Cas9