生物技术通报2017，Vol.33Issue(11)：180-187,8.DOI:10.13560/j.cnki.biotech.bull.1985.2017-0394
应用CRISPR/Cas9系统下调长链非编码RNA HOTAIR
Knockdown of Long Non-coding RNA HOTAIR by CRISPR/Cas9 System
摘要
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,resuhs 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.关键词
CRISPR/Cas9/pgRNA/长链非编码RNA/HOTAIR/增殖/迁移/肿瘤抑制因子Key words
CRISPR/Cas9/pgRNA/LncRNA/HOTAIR/proliferation/migration/tumor suppressor引用本文复制引用
刘宏博,郑鹏,印泽,张同存..应用CRISPR/Cas9系统下调长链非编码RNA HOTAIR[J].生物技术通报,2017,33(11):180-187,8.基金项目
国家自然科学基金项目(31500622),武汉科技大学计划项目(25011401) （31500622）
