title: Synthetic RNA-protein modules integrated with native translation mechanisms to control gene expression in malaria parasites.
abstract: 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.
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139 Massachusetts, USA.
authors.forename: Suresh M
authors.initials: SM
authors.lastname: Ganesan
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139 Massachusetts, USA.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Falla
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139 Massachusetts, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Goldfless
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139 Massachusetts, USA.
authors.forename: Armiyaw S
authors.initials: AS
authors.lastname: Nasamu
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139 Massachusetts, USA.
authors.forename: Jacquin C
authors.initials: JC
authors.lastname: Niles
mesh_terms: D003001:Cloning, Molecular; D005786:Gene Expression Regulation / Q000502:physiology*; D010963:Plasmodium falciparum / Q000235:genetics / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins