Knockdown of Long Non-coding RNA HOTAIR by CRISPR/Cas9 System

doi:10.13560/j.cnki.biotech.bull.1985.2017-0394

Abstract

Abstract: Recently,as a mature gene editing technique,the CRISPR/Cas9 system has been widely applied. However,it is rarely used in the studies of long non-coding RNAs（lncRNAs）,owe to their specific secondary structure. The lncRNA HOTAIR was called oncogenic lncRNA overexpressing in the vast majority of cancer types,regulating various biological processes such as cell growth,apoptosis,proliferation and metastasis. Our study performed that HOTAIR was knocked out using the Cas9 system based on the stably genetic cell lines HeLa-cas9 and a pgRNA library was built. The qRT-PCR analysis suggested that the RNA expression of HOTAIR decreased by 60% with pgRNA/Cas9. MTT proliferation detection and cell scratch test revealed that HeLa cell proliferation and migration were significantly inhibited after the knockout of HOTAIR. Meanwhile,results indicated that it was associated between HOTAIR and some tumor suppressors by measuring the RNA level of p21,p53,pRB and DLC1 after the knockout of HOTAIR. Thus,it provided a novel insight to comprehensively understand the function of HOTAIR in cancer cells.

Key words: CRISPR/Cas9, pgRNA, LncRNA, HOTAIR, proliferation, migration, tumor suppressor

LIU Hong-bo, ZHENG Peng, YIN Ze, ZHANG Tong-cun. Knockdown of Long Non-coding RNA HOTAIR by CRISPR/Cas9 System[J]. Biotechnology Bulletin, 2017, 33(11): 180-187.

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