Premium
Bidirectional Regulation of mRNA Translation in Mammalian Cells by Using PUF Domains
Author(s) -
Cao Jicong,
Arha Manish,
Sudrik Chaitanya,
Schaffer David V.,
Kane Ravi S.
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201402095
Subject(s) - translation (biology) , messenger rna , downregulation and upregulation , repressor , mutant , synthetic biology , rna , microbiology and biotechnology , rna binding protein , translational regulation , biology , computational biology , protein biosynthesis , gene expression , gene , chemistry , genetics
The regulation of gene expression is crucial in diverse areas of biological science, engineering, and medicine. A genetically encoded system based on the RNA binding domain of the Pumilio and FBF (PUF) proteins was developed for the bidirectional regulation (i.e., either upregulation or downregulation) of the translation of a target mRNA. PUF domains serve as designable scaffolds for the recognition of specific RNA elements and the specificity can be easily altered to target any 8‐nucleotide RNA sequence. The expression of a reporter could be varied by over 17‐fold when using PUF‐based activators and repressors. The specificity of the method was established by using wild‐type and mutant PUF domains. Furthermore, this method could be used to activate the translation of target mRNA downstream of PUF binding sites in a light‐dependent manner. Such specific bidirectional control of mRNA translation could be particularly useful in the fields of synthetic biology, developmental biology, and metabolic engineering.