Sequence-specific RNA Photocleavage by Single-stranded DNA in Presence of Riboflavin.
Sequence-specific RNA Photocleavage by Single-stranded DNA in Presence of Riboflavin.
复制标题
核黄素存在下单链 DNA 的序列特异性 RNA 光裂解
DOI：
10.1038/srep15039
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发表时间：
2015-10-13
影响因子：
4.6
通讯作者：
Tang Z
中科院分区：
文献类型：
--
作者：
Zhao Y;Chen G;Yuan Y;Li N;Dong J;Huang X;Cui X;Tang Z
人们一直在努力开发新的方法来实现序列特异性RNA降解，这可能会导致靶基因的表达抑制。本文报道了一种利用未修饰的短DNA寡核苷酸进行序列识别，以内源性小分子维生素B2（核黄素）为光敏剂，实现序列特异性光切割靶RNA的简单方法。DNA链与靶序列互补，形成DNA/RNA双链体，中间含有G·U摆动。切割反应在双链体中G·U摆动的U下游的核苷处通过氧化消除机制得到非天然RNA末端，整个过程处于以光为开关的严格控制之下，这意味着切割可以根据特定的空间和时间要求进行。这种方法的生物相容性使得DNA链与核黄素的组合成为RNA操作的有前途的分子工具。
Constant efforts have been made to develop new method to realize sequence-specific RNA degradation, which could cause inhibition of the expression of targeted gene. Herein, by using an unmodified short DNA oligonucleotide for sequence recognition and endogenic small molecue, vitamin B2 (riboflavin) as photosensitizer, we report a simple strategy to realize the sequence-specific photocleavage of targeted RNA. The DNA strand is complimentary to the target sequence to form DNA/RNA duplex containing a G•U wobble in the middle. The cleavage reaction goes through oxidative elimination mechanism at the nucleoside downstream of U of the G•U wobble in duplex to obtain unnatural RNA terminal and the whole process is under tight control by using light as switch, which means the cleavage could be carried out according to specific spatial and temporal requirements. The biocompatibility of this method makes the DNA strand in combination with riboflavin a promising molecular tool for RNA manipulation.