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微RNA聚焦的CRISPR/Cas9屏幕识别了miR-142作为爱斯坦-巴尔病毒重新激活的关键调节者

Yan Chen¹, Rodney P Kincaid¹, Kelley Bastin¹

¹Vaccine and Gene Therapy Institute, Oregon Health & Science University, Beaverton, Oregon, United States of America.

|June 17, 2024

概括

微RNA-142 (miR-142) 限制了埃普斯坦-巴尔病毒 (EBV) 从潜伏期的重新激活. 抑制miR-142可以通过调节B细胞受体信号通路来增强EBV的性基因表达.

科学领域:

病毒学病毒学
分子生物学分子生物学
免疫学免疫学免疫学

背景情况:

埃普斯坦-巴尔病毒 (EBV) 通过潜伏期建立终身持久感染.
从潜伏阶段到EBV溶解阶段的重新激活对于病毒持久性至关重要,但人们对其了解甚少.
病毒和细胞微RNAs (miRNAs) 在EBV感染和发病过程中发挥作用.

研究的目的:

为了识别宿主细胞的miRNAs,这些miRNAs调节了从EBV延迟转变到溶液周期的过渡.
阐明这些miRNAs控制EBV重新激活的机制.

主要方法:

在EBV阳性伯基特淋巴瘤细胞中基于CRISPR/Cas9的基因选.
通过与抗Ig抗体交联的B细胞受体 (BCR) 诱导EBV的活性化.
在Ago2-PAR-CLIP实验中,用于识别miRNA目标.
信号通路的药理抑制 (Mek, Raf).

主要成果:

确定miR-142是限制EBV重新激活的关键调节剂.
基因切除miR-142增加了立即早期和早期的lytic基因表达.
miR-142针对BCR下游的Erk/MAPK信号通路的组件.
破坏miR-142增强SOS1水平和Mek酸化,这些被Mek/Raf抑制剂所拯救.

结论:

miR-142 作为一种限制宿主因子,限制了EBV 溶解周期的进入.
miR-142调节由BCR激活启动的SOS1/Ras/Raf/Mek/Erk信号级联.
了解miR-142的作用为控制EBV的重新激活和发病提供了洞察力.

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MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...