title: Engineering ribosomal leaky scanning and upstream open reading frames for precise control of protein translation.
abstract: We have employed upstream open reading frames (uORFs) to systematically tune the translation levels of recombinant proteins. We present the design principles that guided the development of this technology and provide information that may help others in implementing synthetic uORFs for their own applications. We also report on recent applications to our own research projects, including the coupling of uORF and translation initiation site (TIS) engineering with small molecule-inducible post-translational control. Finally, we discuss opportunities to investigate and potentially engineer gene-specific translational responses to cellular stress.
authors.affiliation: Department of Chemical Engineering; Stanford University; Stanford, CA USA.
authors.forename: Joshua P
authors.initials: JP
authors.lastname: Ferreira
authors.affiliation: Department of Chemical Engineering; Stanford University; Stanford, CA USA.
authors.forename: William L
authors.initials: WL
authors.lastname: Noderer
authors.affiliation: Department of Chemical Engineering; Stanford University; Stanford, CA USA.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Diaz de Arce
authors.affiliation: Department of Chemical Engineering; Stanford University; Stanford, CA USA.
authors.forename: Clifford L
authors.initials: CL
authors.lastname: Wang
mesh_terms: D051701:Activating Transcription Factor 4 / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010442:Peptide Chain Initiation, Translational; D011401:Promoter Regions, Genetic / Q000187:drug effects; D016283:Proto-Oncogene Proteins p21(ras) / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012270:Ribosomes / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014295:Trimethoprim / Q000494:pharmacology
chemical_list: C497109:ATF4 protein, human; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D051701:Activating Transcription Factor 4; D049452:Green Fluorescent Proteins; D014295:Trimethoprim; D016283:Proto-Oncogene Proteins p21(ras)
keywords: cellular stress; post-transcriptional regulation; ribosome scanning; translation initiation; uORF; upstream open reading frames