Orthogonal Protein-Responsive mRNA Switches for Mammalian Synthetic Biology.,ACS Synthetic Biology

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Orthogonal Protein-Responsive mRNA Switches for Mammalian Synthetic Biology.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2019-12-13 , DOI: 10.1021/acssynbio.9b00343
Hiroki Ono _{1,

2} , Shunsuke Kawasaki ₁ , Hirohide Saito ₁

Affiliation

The lack of available genetic modules is a fundamental issue in mammalian synthetic biology. Especially, the variety of genetic parts for translational control are limited. Here we report a new set of synthetic mRNA-based translational switches by engineering RNA-binding proteins (RBPs) and RBP-binding RNA motifs (aptamers) that perform strong translational repression. We redesigned the RNA motifs with RNA scaffolds and improved the efficiency of the repression to target RBPs. Using new and previously reported mRNA switches, we demonstrated that the orthogonality of translational regulation was ensured among five different RBP-responsive switches. Moreover, the new switches functioned not only with plasmid introduction, but also with RNA-only delivery, which provides a transient and safer regulation of expression. The translational regulators using RNA-protein interactions provide an alternative strategy to construct complex genetic circuits for future cell engineering and therapeutics.

中文翻译：

哺乳动物合成生物学的正交蛋白质响应mRNA开关。

缺乏可用的遗传模块是哺乳动物合成生物学中的一个基本问题。特别地，用于翻译控制的遗传部分的种类是有限的。在这里，我们通过工程RNA结合蛋白（RBPs）和RBP结合RNA基序（适体）报告了一组新的基于mRNA的合成翻译开关，它们执行了强大的翻译抑制作用。我们用RNA支架重新设计了RNA模体，并提高了抑制靶向RBP的效率。使用新的和以前报道的mRNA开关，我们证明了翻译调控的正交性在五个不同的RBP响应开关之间得到了保证。此外，新的开关不仅通过质粒导入起作用，而且还通过仅RNA递送起作用，从而提供了瞬时且安全的表达调控。

更新日期：2019-12-17

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