Until recently, RNA-RNA interactions were mainly identified by crosslinking RNAs with interacting proteins, RNA proximity ligation and deep sequencing. Recently, AMT-based direct RNA crosslinking was established. Yet, several steps of these procedures are rather inefficient, reducing the output of identified interaction partners. To increase the local concentration of RNA ends, interacting RNAs are often fragmented. However, the resulting 2',3'-cyclic phosphate and 5'-OH ends are not accepted by T4 RNA ligase and have to be converted to 3'-OH and 5'-phosphate ends. Using an artificial mRNA/sRNA pair, we optimized the workflow downstream of the crosslinking reaction in vitro. The use of a tRNA ligase allows direct fusion of 2',3'-cyclic phosphate and 5'-OH RNA ends.

中文翻译：

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使用体外AMT交联检测mRNA-sRNA相互作用的简化方案。

直到最近，RNA-RNA的相互作用主要是通过将RNA与相互作用的蛋白质交联，RNA邻近连接和深度测序来鉴定的。最近，建立了基于AMT的直接RNA交联。然而，这些程序的几个步骤效率很低，从而降低了确定的交互伙伴的输出。为了增加RNA末端的局部浓度，相互作用的RNA通常被片段化。但是，所得的2'，3'-环状磷酸酯和5'-OH末端未被T4 RNA连接酶接受，必须将其转化为3'-OH和5'-磷酸酯末端。使用人工的mRNA /的sRNA对，我们优化了交联反应的下游工作流体外。tRNA连接酶的使用允许2'，3'-环状磷酸酯和5'-OH RNA末端直接融合。