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基因组规模激活屏幕识别了调节基因邻近的 lncRNA 位点

Julia Joung^1,2,3,4, Jesse M Engreitz², Silvana Konermann^2,3,4

¹Department of Biological Engineering, MIT, Cambridge, Massachusetts 02139, USA.

|August 10, 2017

概括

研究人员发现了11个长非编码RNA (lncRNA) 位点,在黑色素瘤中赋予BRAF抑制剂耐药性. 激活这些lncRNA基因可以局部调节附近的蛋白质编码基因,从而提供新的治疗点.

科学领域:

基因组学
分子生物学
癌症研究

背景情况:

哺乳动物基因组包含许多参与细胞过程的长非编码RNA (lncRNA) 位点.
虽然一些lncRNAs在trans中起作用,但许多人则在cis中调节附近的基因.
识别功能性 lncRNA 位点及其机制仍然具有挑战性.

研究的目的:

开发基因组规模的CRISPR-Cas9激活屏幕以识别功能IncRNA位置.
发现影响特定表型的非编码位点,如耐药性.

主要方法:

一个全基因组的CRISPR-Cas9激活屏幕,针对超过10,000个 lncRNA转录起点.
在 lncRNA 激活时对黑色素瘤细胞进行表型分析.
候选 lncRNA 位点的详细表征,包括 EMICERI.

主要成果:

在人类黑色素瘤细胞中发现了11个 lncRNA 位点,这些位点介导了对 BRAF 抑制剂的抵抗.
大多数已识别的 lncRNA 位置似乎调节了附近基因的表达.
转录激活EMICERI导致四个邻近的蛋白质编码基因的剂量依赖激活,从而产生耐药性.

结论:

开发的CRISPR-Cas9激活屏幕对于发现非编码位点的功能是有效的.
在调节基因表达和细胞功能,包括药物耐药性方面, lncRNAs 发挥着重要作用.
这种方法为非编码位点的系统功能基因组学提供了一个工具包.

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