title: Site-specific promoter caging enables optochemical gene activation in cells and animals.
abstract: In cell and molecular biology, double-stranded circular DNA constructs, known as plasmids, are extensively used to express a gene of interest. These gene expression systems rely on a specific promoter region to drive the transcription of genes either constitutively (i.e., in a continually "ON" state) or conditionally (i.e., in response to a specific transcription initiator). However, controlling plasmid-based expression with high spatial and temporal resolution in cellular environments and in multicellular organisms remains challenging. To overcome this limitation, we have site-specifically installed nucleobase-caging groups within a plasmid promoter region to enable optochemical control of transcription and, thus, gene expression, via photolysis of the caging groups. Through the light-responsive modification of plasmid-based gene expression systems, we have demonstrated optochemical activation of an exogenous fluorescent reporter gene in both tissue culture and a live animal model, as well as light-induced overexpression of an endogenous signaling protein.
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina 27695, United States.
authors.forename: James
authors.initials: J
authors.lastname: Hemphill
authors.forename: Jeane
authors.initials: J
authors.lastname: Govan
authors.forename: Rajendra
authors.initials: R
authors.lastname: Uprety
authors.forename: Michael
authors.initials: M
authors.lastname: Tsang
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D000818:Animals; D001483:Base Sequence; D004270:DNA, Circular / Q000737:chemistry / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D010782:Photolysis; D010957:Plasmids / Q000737:chemistry / Q000235:genetics*; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D015533:Transcriptional Activation; D025521:Tumor Suppressor Proteins; D014466:Ultraviolet Rays; D015027:Zebrafish; D000097868:Polo-like Kinases
chemical_list: D004270:DNA, Circular; D049452:Green Fluorescent Proteins; D017346:Protein Serine-Threonine Kinases; D025521:Tumor Suppressor Proteins; C485184:enhanced green fluorescent protein; C486256:PLK3 protein, human; D000097868:Polo-like Kinases