Targeted RNA m6A Editing Using Engineered CRISPR-Cas9 Conjugates.
Targeted RNA m6A Editing Using Engineered CRISPR-Cas9 Conjugates.
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DOI：
10.1007/978-1-0716-1374-0_23
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发表时间：
2021
影响因子：
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通讯作者：
Xiao-Min Liu;Shu-Bing Qian
中科院分区：
文献类型：
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作者：
Xiao-Min Liu;Shu-Bing Qian
N6-甲基腺苷(M6A)是一种重要的表位转录标记，在RNA代谢中发挥着重要的作用，包括RNA的稳定性、mRNA的翻译和RNA的结构重排。不同RNA区域的M6A修饰可能具有不同的分子效应。在这里，我们描述了一种基于CRISPR-Cas9的方法，该方法能够在不改变核苷酸序列的情况下对内源RNA分子进行有针对性的m6A添加或移除。通过将催化失活的Cas9与工程的m6A修饰酶融合，可编程的m6A编辑能够在所需的位置实现RNA甲基化和去甲基化，促进对m6A的区域效应的剖析，并使RNA操作工具箱多样化。
N6-methyladenosine (m6A) is a major epitranscriptomic mark exerting crucial diverse roles in RNA metabolisms, including RNA stability, mRNA translation, and RNA structural rearrangement. m6A modifications at different RNA regions may have distinct molecular effects. Here, we describe a CRISPR-Cas9-based approach that enables targeted m6A addition or removal on endogenous RNA molecules without altering the nucleotide sequence. By fusing a catalytically inactive Cas9 with engineered m6A modification enzymes, the programmable m6A editors are capable of achieving RNA methylation and demethylation at desired sites, facilitating dissection of regional effects of m6A and diversifying the toolkits for RNA manipulation.