Caliciviral protein-based artificial translational activator for mammalian gene circuits with RNA-only delivery
Caliciviral protein-based artificial translational activator for mammalian gene circuits with RNA-only delivery
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DOI：
10.1038/s41467-020-15061-x
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发表时间：
2020-03-10
影响因子：
16.6
通讯作者：
Saito, Hirohide
中科院分区：
文献类型：
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作者：
Nakanishi, Hideyuki;Saito, Hirohide
基于合成RNA的基因电路可实现复杂的基因调节，而无需插入诱变的风险。虽然已经使用了各种RNA结合蛋白在基因回路中进行翻译抑制，但尚未实现合成mRNA的直接翻译激活。在这里，我们开发了基于Caliciviral VPG的转化激活剂（CAVT），该转化剂（CAVT）激活了没有规范5'CAP的合成mRNA的翻译。可以通过更改目标mRNA中CAVT结合基序的位置，序列和修饰的核苷来调节翻译水平，从而使不同mRNA的同时翻译激活和仅RNA递送的不同mRNA抑制。我们通过用CAVT调整翻译水平来证明对凋亡和基因组编辑的有效调节。此外，我们设计可编程的CAVT，对内源性microRNA或小分子做出反应，从而从合成mRNA中实现了细胞状态特异性和条件翻译激活。 CAVT将成为生物学研究和未来治疗应用的合成生物学的重要工具。
Synthetic RNA-based gene circuits enable sophisticated gene regulation without the risk of insertional mutagenesis. While various RNA binding proteins have been used for translational repression in gene circuits, the direct translational activation of synthetic mRNAs has not been achieved. Here we develop Caliciviral VPg-based Translational activator (CaVT), which activates the translation of synthetic mRNAs without the canonical 5'-cap. The level of translation can be modulated by changing the locations, sequences, and modified nucleosides of CaVT-binding motifs in the target mRNAs, enabling the simultaneous translational activation and repression of different mRNAs with RNA-only delivery. We demonstrate the efficient regulation of apoptosis and genome editing by tuning translation levels with CaVT. In addition, we design programmable CaVT that responds to endogenous microRNAs or small molecules, achieving both cell-state-specific and conditional translational activation from synthetic mRNAs. CaVT will become an important tool in synthetic biology for both biological studies and future therapeutic applications.