Earth-Life Science Institute, Tokyo Institute of Technology, Ookayama, Meguro-ku, Tokyo 152-8550, Japan.
School of Life Science and Technology, Tokyo Institute of Technology, Meguro-ku, Tokyo 152-8550, Japan.
Biomacromolecules. 2024 Jan 8;25(1):355-365. doi: 10.1021/acs.biomac.3c01024. Epub 2023 Dec 5.
RNA-binding proteins participate in diverse cellular processes, including DNA repair, post-transcriptional modification, and cancer progression through their interactions with RNAs, making them attractive for biotechnological applications. While nature provides an array of naturally occurring RNA-binding proteins, developing RNA-binding peptides remains challenging. In particular, tailoring peptides to target single-stranded RNA with low complexity is difficult due to the inherent structural flexibility of RNA molecules. Here, we developed a codon-restricted mRNA display and identified multiple peptides from a peptide library that bind to poly(C) and poly(A) RNA with s ranging from micromolar to submicromolar concentrations. One of the newly identified peptides is capable of binding to the cytosine-rich sequences of the oncogenic Cdk6 3'UTR RNA and lncRNA, with affinity comparable to that of the endogenous binding protein. Hence, we present a novel platform for discovering single-stranded RNA-binding peptides that offer promising avenues for regulating RNA functions.
RNA 结合蛋白通过与 RNA 的相互作用参与多种细胞过程，包括 DNA 修复、转录后修饰和癌症进展，因此它们成为生物技术应用的有吸引力的目标。虽然自然界提供了一系列天然存在的 RNA 结合蛋白，但开发 RNA 结合肽仍然具有挑战性。特别是，由于 RNA 分子固有的结构灵活性，针对具有低复杂度的单链 RNA 定制肽非常困难。在这里，我们开发了一种密码子受限的 mRNA 展示，并从肽文库中鉴定出多个能够以微摩尔至亚微摩尔浓度范围与聚（C）和聚（A）RNA 结合的肽。新鉴定出的肽之一能够与致癌 Cdk6 3'UTR RNA 和 lncRNA 的富含胞嘧啶序列结合，亲和力可与内源性结合蛋白相媲美。因此，我们提出了一种发现单链 RNA 结合肽的新平台，为调节 RNA 功能提供了有前途的途径。
