通过AGO2 RNA免疫沉淀和荧光素酶筛选试验鉴定miRNA靶标的环状RNA
Identification of circRNAs for miRNA Targets by Argonaute2 RNA Immunoprecipitation and Luciferase Screening Assays.
作者信息
Li Yan, Chen Bing, Huang Shenglin
机构信息
Fudan University Shanghai Cancer Center, Institutes of Biomedical Sciences and Shanghai Medical College, Fudan University, Shanghai, China.
出版信息
Methods Mol Biol. 2018;1724:209-218. doi: 10.1007/978-1-4939-7562-4_17.
Circular RNAs from back-spliced exons (circRNAs) represent a novel class of widespread and endogenous RNAs in eukaryotes. circRNAs may bind to microRNAs (miRNAs) and inhibit the activity of miRNAs. Alternatively, miRNAs could also directly target circRNAs and regulate the expression of circRNAs. Here we describe the Argonaute2 (AGO2) RNA immunoprecipitation (RIP) and luciferase screening assays to identify the interaction between circRNAs and miRNAs. The AGO2 RIP assay evaluates the potential of the interaction between circRNAs and miRNAs. The luciferase screening assay investigates the targeting miRNAs and the detail binding sites for a specific circRNA.
来自反向剪接外显子的环状RNA（circRNAs）是真核生物中一类新型的广泛存在的内源性RNA。circRNAs可能与微小RNA（miRNAs）结合并抑制miRNAs的活性。或者，miRNAs也可以直接靶向circRNAs并调节circRNAs的表达。在这里，我们描述了AGO2 RNA免疫沉淀（RIP）和荧光素酶筛选试验，以鉴定circRNAs与miRNAs之间的相互作用。AGO2 RIP试验评估circRNAs与miRNAs之间相互作用的可能性。荧光素酶筛选试验研究靶向miRNAs以及特定circRNA的详细结合位点。
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