Exonuclease assisted mapping of protein-RNA interactions (ePRINT)
Exonuclease assisted mapping of protein-RNA interactions (ePRINT)
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DOI：
10.1101/2023.05.16.540978
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发表时间：
2023-05
期刊：
影响因子：
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通讯作者：
Sophie Hawkins;Alexandre Mondaini;Seema Namboori;Asif Javed;A. Bhinge
中科院分区：
文献类型：
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作者：
Sophie Hawkins;Alexandre Mondaini;Seema Namboori;Asif Javed;A. Bhinge
RNA加工是基因调控的基本模式，在包括癌症和神经退行性疾病在内的多种疾病中受到干扰。RNA结合蛋白（RBP）通过物理结合RNA分子来调节RNA加工的关键方面，包括选择性剪接、mRNA降解和定位。目前绘制这些相互作用的方法，如CLIP，依赖于一次纯化单个蛋白质。我们已经开发了一种新的方法（ePRINT）来在全球范围内绘制RBP-RNA相互作用网络，而无需纯化单个RBPs。ePRINT可以精确定位RBP结合位点的5'端，并可以发现感兴趣RBP的直接和间接靶点。重要的是，ePRINT还可以发现在细胞命运转变之间差异激活的RBP，例如，当神经祖细胞分化为神经元时。鉴于其多功能性，ePRINT作为发育、健康和疾病中RNA调控的研究工具具有巨大的应用潜力。
RNA processing is a fundamental mode of gene regulation that is perturbed in a variety of diseases including cancer and neurodegenerative disorders. RNA-binding proteins (RBPs) regulate key aspects of RNA processing including alternative splicing, mRNA degradation and localization by physically binding RNA molecules. Current methods to map these interactions, such as CLIP, rely on purifying single proteins at a time. We have developed a new method (ePRINT) to map RBP-RNA interaction networks on a global scale without purifying individual RBPs. ePRINT allows precise mapping of the 5’ end of the RBP binding site, and can uncover direct and indirect targets of an RBP of interest. Importantly, ePRINT can also uncover RBPs that are differentially activated between cell fate transitions, for instance, as neural progenitors differentiate into neurons. Given its versatility, ePRINT has vast application potential as an investigative tool for RNA regulation in development, health and disease.