Synthetic RNA-protein modules integrated with native translation mechanisms to control gene expression in malaria parasites
Synthetic RNA-protein modules integrated with native translation mechanisms to control gene expression in malaria parasites
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DOI：
10.1038/ncomms10727
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发表时间：
2016-03-01
影响因子：
16.6
通讯作者：
Niles, Jacquin C.
中科院分区：
文献类型：
--
作者：
Ganesan, Suresh M.;Falla, Alejandra;Niles, Jacquin C.
基因表达的合成转录后调控对于理解基础生物学和在合成生物学中编程新的细胞过程是重要的。以前用于调节真核生物翻译的策略集中在破坏翻译中的各个步骤，包括起始和mRNA切割。在强调模块化和跨有机体的功能，这些系统被设计成正交运行本地控制机制。在这里，我们介绍了一个广泛适用的策略，通过整合合成的翻译控制通过小分子调节的RNA蛋白模块与天然机制，同时调节细胞RNA命运的多个方面，有力地控制蛋白质翻译。我们证明，这种策略减少了“泄漏”，以提高整体表达的动态范围，并可以实现而不牺牲模块化和跨有机体的功能。我们说明这在酿酒酵母和非模型人类疟疾寄生虫，恶性疟原虫。鉴于有限的功能遗传学工具包可用于恶性疟原虫，我们建立了这种策略的实用性定义必需基因。
Synthetic posttranscriptional regulation of gene expression is important for understanding fundamental biology and programming new cellular processes in synthetic biology. Previous strategies for regulating translation in eukaryotes have focused on disrupting individual steps in translation, including initiation and mRNA cleavage. In emphasizing modularity and crossorganism functionality, these systems are designed to operate orthogonally to native control mechanisms. Here we introduce a broadly applicable strategy for robustly controlling protein translation by integrating synthetic translational control via a small-molecule-regulated RNA-protein module with native mechanisms that simultaneously regulate multiple facets of cellular RNA fate. We demonstrate that this strategy reduces 'leakiness' to improve overall expression dynamic range, and can be implemented without sacrificing modularity and cross-organism functionality. We illustrate this in Saccharomyces cerevisae and the non-model human malarial parasite, Plasmodium falciparum. Given the limited functional genetics toolkit available for P. falciparum, we establish the utility of this strategy for defining essential genes.