Orthogonal Protein-Responsive mRNA Switches for Mammalian Synthetic Biology
Orthogonal Protein-Responsive mRNA Switches for Mammalian Synthetic Biology
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DOI：
10.1021/acssynbio.9b00343
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发表时间：
2020-01-01
影响因子：
4.7
通讯作者：
Saito, Hirohide
中科院分区：
文献类型：
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作者：
Ono, Hiroki;Kawasaki, Shunsuke;Saito, Hirohide
缺乏可用的遗传模块是哺乳动物合成生物学的一个基本问题。特别是，用于翻译控制的遗传部分的多样性是有限的。在这里，我们报告了一组新的基于 mRNA 的合成翻译开关，通过设计 RNA 结合蛋白 (RBP) 和 RBP 结合 RNA 基序（适体）来执行强翻译抑制。我们用 RNA 支架重新设计了 RNA 基序，并提高了对目标 RBP 的抑制效率。使用新的和先前报道的 mRNA 开关，我们证明了五种不同的 RBP 响应开关之间翻译调控的正交性得到了保证。此外，新的开关不仅可以用于质粒导入，还可以用于仅 RNA 的传递，从而提供瞬时且更安全的表达调节。使用 RNA 蛋白质相互作用的翻译调节因子为未来的细胞工程和治疗提供了构建复杂遗传电路的替代策略。
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.