Synthetic RNA–protein modules integrated with native translation mechanisms to control gene expression in malaria parasites

• Published in: Nature communications Vol. 7; no. 1; p. 10727
• Main Authors: Ganesan, Suresh M., Falla, Alejandra, Goldfless, Stephen J., Nasamu, Armiyaw S., Niles, Jacquin C.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.03.2016; Nature Publishing Group; Nature Portfolio
• Subjects: 13/109; 13/31; 13/44; 38/1; 38/35; 38/77; 631/208/199; 631/326/417/2550; 631/337/458; 631/61/338/552; 82/51; Cloning, Molecular; Gene Expression Regulation - physiology; Humanities and Social Sciences; multidisciplinary; Plasmodium falciparum - genetics; Plasmodium falciparum - metabolism; Recombinant Proteins; RNA Processing, Post-Transcriptional; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Science; Science (multidisciplinary)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms10727

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 cross-organism 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. Current strategies for regulatory control of gene expression are orthogonal to the host organism mechanisms. Here the authors demonstrate an RNA aptamer controlled system integrated into native regulatory pathways in the parasite Plasmodium falciparum .