title: Bidirectional regulation of mRNA translation in mammalian cells by using PUF domains.
abstract: 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.
authors.affiliation: Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180 (USA).
authors.forename: Jicong
authors.initials: J
authors.lastname: Cao
authors.forename: Manish
authors.initials: M
authors.lastname: Arha
authors.forename: Chaitanya
authors.initials: C
authors.lastname: Sudrik
authors.forename: David V
authors.initials: DV
authors.lastname: Schaffer
authors.forename: Ravi S
authors.initials: RS
authors.lastname: Kane
mesh_terms: D000818:Animals; D005786:Gene Expression Regulation; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D012333:RNA, Messenger
keywords: RNA; gene expression; gene technology; inducible gene expression; optogenetics