人工ＲＮＡを活用した高効率・汎用的な細胞リプログラミング技術の確立

利用人工RNA建立高效、多功能的细胞重编程技术

• 批准号: 17F17390
• 负责人: 齊藤 博英
• 金额: $ 1.54万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2017
• 资助国家: 日本
• 起止时间: 2017-11-10 至 2020-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17F17390/
• 关键词: RNA methylation; microRNA; iPSC differentiation

In this project, we first demonstrated that by a simple methylation of pseudouridine can significantly alter the thermodynamic properties of RNAs, which results in the enhancement of mRNA expression rate, and ambiguously influence the binding between microRNAs with their complementary RNA sequences.We have then use T7 polymerase based in vitro RNA synthesis to produce mRNAs containing various types of methylated nucleotide, and examined their performance in a variety of cells, including iPSC and differentiated iPSCs. We found that on all cell types tested, including iPSCs, the mRNAs containing methylated pseudouridine exhibited significantly higher protein production rate, indicating the impact of methylation of RNA is conserved among cell types. However, the fold difference in protein expression between native and methylated mRNAs are different for each cell types, suggesting each cell response to RNA methylation in different ways.

在本项目中，我们首先证明了通过一个简单的假尿苷甲基化可以显著改变RNA的热力学性质，从而导致mRNA表达率的提高，并模糊地影响microRNA与其互补RNA序列之间的结合，然后我们使用基于T7聚合酶的体外RNA合成来产生含有各种类型的甲基化核苷酸的mRNA，并检查了它们在各种细胞中的表现，包括iPSC和分化的iPSC。我们发现，在所有测试的细胞类型中，包括iPSC，含有甲基化假尿苷的mRNA表现出显著更高的蛋白质产生率，表明RNA甲基化的影响在细胞类型中是保守的。然而，对于每种细胞类型，天然mRNA和甲基化mRNA之间的蛋白质表达的倍数差异是不同的，这表明每种细胞以不同的方式对RNA甲基化做出反应。

项目成果

N-1-Methyl-pseudouridine to enhance the performance of RNA tools in cells

N-1-甲基-伪尿苷可增强细胞中 RNA 工具的性能

• 发表时间: 2018
• 作者: Yi Kuang;Callum J.C. Parr;Shunsuke Wada;Kaoru R. Komatsu;Hirohito Saito
• 通讯作者: Hirohito Saito