Argonaute binding within human nuclear RNA and its impact on alternative splicing.
Argonaute binding within human nuclear RNA and its impact on alternative splicing.
复制标题
人核RNA中的Argonaute结合及其对选择性剪接的影响。
DOI：
10.1261/rna.078707.121
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发表时间：
2021-09
期刊：
影响因子：
--
通讯作者：
Corey DR
中科院分区：
文献类型：
--
作者：
Chu Y;Yokota S;Liu J;Kilikevicius A;Johnson KC;Corey DR
哺乳动物RNA干扰（RNAi）通常与细胞质中基因表达的调控有关。然而，合成RNA也可以通过RNAi途径来调节转录和剪接。虽然通过合成RNA进行的核调控可能是稳健的，但一个关键的未回答的问题是核RNAi的内源性功能是否存在于哺乳动物细胞中。我们使用增强型交联免疫沉淀（eCLIP）结合RNA测序（RNA-seq）和多个AGO敲除细胞系，绘制了核RNA内的AGO 2蛋白结合位点。最强的AGO 2结合位点被定位到微小RNA（miRNAs）。最丰富的miRNAs在细胞质和细胞核之间分布相似，没有证据表明促进miRNAs在一个区室中定位的机制。除了miRNAs之外，大多数统计学上显著的AGO 2结合在内含子内。剪接变化通过RT-PCR证实，并通过与AGO 2结合位点互补的合成miRNA模拟物重现。这些数据支持miRNAs可以控制基因剪接的假设。虽然核RNAi蛋白有可能成为天然的调节机制，但需要仔细研究以确定正常生理和疾病的关键RNA驱动因素。
Mammalian RNA interference (RNAi) is often linked to the regulation of gene expression in the cytoplasm. Synthetic RNAs, however, can also act through the RNAi pathway to regulate transcription and splicing. While nuclear regulation by synthetic RNAs can be robust, a critical unanswered question is whether endogenous functions for nuclear RNAi exist in mammalian cells. Using enhanced crosslinking immunoprecipitation (eCLIP) in combination with RNA sequencing (RNA-seq) and multiple AGO knockout cell lines, we mapped AGO2 protein binding sites within nuclear RNA. The strongest AGO2 binding sites were mapped to micro RNAs (miRNAs). The most abundant miRNAs were distributed similarly between the cytoplasm and nucleus, providing no evidence for mechanisms that facilitate localization of miRNAs in one compartment versus the other. Beyond miRNAs, most statistically significant AGO2 binding was within introns. Splicing changes were confirmed by RT-PCR and recapitulated by synthetic miRNA mimics complementary to the sites of AGO2 binding. These data support the hypothesis that miRNAs can control gene splicing. While nuclear RNAi proteins have the potential to be natural regulatory mechanisms, careful study will be necessary to identify critical RNA drivers of normal physiology and disease.
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影响因子：
64.5
作者：
Hafner M;Landthaler M;Burger L;Khorshid M;Hausser J;Berninger P;Rothballer A;Ascano M Jr;Jungkamp AC;Munschauer M;Ulrich A;Wardle GS;Dewell S;Zavolan M;Tuschl T
通讯作者：
Tuschl T
DOI：
10.1073/pnas.0808830105
发表时间：
2008-10-21
影响因子：
11.1
作者：
Kim, Daniel H.;Saetrom, Pal;Rossi, John J.
通讯作者：
Rossi, John J.
影响因子：
4.5
作者：
Jeffries, Clark D.;Fried, Howard M.;Perkins, Diana O.
通讯作者：
Perkins, Diana O.
影响因子：
4
作者：
Liu, Zhongtian;Johnson, Samantha T.;Corey, David R.
通讯作者：
Corey, David R.
影响因子：
64.8
作者：
Ghandi, Mahmoud;Huang, Franklin W.;Sellers, William R.
通讯作者：
Sellers, William R.