title: Optochemical control of RNA interference in mammalian cells.
abstract: Short interfering RNAs (siRNAs) and microRNAs (miRNAs) have been widely used in mammalian tissue culture and model organisms to selectively silence genes of interest. One limitation of this technology is the lack of precise external control over the gene-silencing event. The use of photocleavable protecting groups installed on nucleobases is a promising strategy to circumvent this limitation, providing high spatial and temporal control over siRNA or miRNA activation. Here, we have designed, synthesized and site-specifically incorporated new photocaged guanosine and uridine RNA phosphoramidites into short RNA duplexes. We demonstrated the applicability of these photocaged siRNAs in the light-regulation of the expression of an exogenous green fluorescent protein reporter gene and an endogenous target gene, the mitosis motor protein, Eg5. Two different approaches were investigated with the caged RNA molecules: the light-regulation of catalytic RNA cleavage by RISC and the light-regulation of seed region recognition. The ability to regulate both functions with light enables the application of this optochemical methodology to a wide range of small regulatory RNA molecules.
authors.affiliation: Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA, Department of Chemistry, College of William & Mary, Williamsburg, VA 32187, USA, Center for Structural Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA and Department of Chemistry, University of Pittsburgh, Pittsburgh, PA 15260, USA.
authors.forename: Jeane M
authors.initials: JM
authors.lastname: Govan
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Young
authors.forename: Hrvoje
authors.initials: H
authors.lastname: Lusic
authors.forename: Qingyang
authors.initials: Q
authors.lastname: Liu
authors.forename: Mark O
authors.initials: MO
authors.lastname: Lively
authors.forename: Alexander
authors.initials: A
authors.lastname: Deiters
mesh_terms: D052117:Benzodioxoles / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D006151:Guanosine / Q000737:chemistry; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D016547:Kinesins / Q000235:genetics; D008027:Light; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000528:radiation effects*; D014529:Uridine / Q000737:chemistry
chemical_list: D052117:Benzodioxoles; C470243:KIF11 protein, human; D009943:Organophosphorus Compounds; D034741:RNA, Small Interfering; C434331:phosphoramidite; D006151:Guanosine; D049452:Green Fluorescent Proteins; D016547:Kinesins; D014529:Uridine
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