title: Construction of a vector generating both siRNA and a fluorescent reporter: a siRNA study in cultured neurons.
abstract: RNA interference is an important tool for gene silencing. However, its application to primary cultured cells has been limited by low transfection efficiencies. In this work we developed a vector which encodes both siRNA and red fluorescent protein. Using this vector we could markedly suppress green fluorescent protein (GFP) and bim an endogenous gene. Primary cultured cortical neurons transfected with siRNA against doublecortin showed that doublecortin expression was significantly inhibited in nearly all the transfected neurons. This vector identifies the transfected cells and should be useful for loss-of-gene function studies in neurons.
authors.affiliation: Department of Anatomy and Cell Biology, University of Ulsan College of Medicine, Seoul 138-736, Korea.
authors.forename: Seung Yong
authors.initials: SY
authors.lastname: Yoon
authors.forename: Jung Eun
authors.initials: JE
authors.lastname: Choi
authors.forename: Onyou
authors.initials: O
authors.lastname: Hwang
authors.forename: Hea Nam
authors.initials: HN
authors.lastname: Hong
authors.forename: Heuiran
authors.initials: H
authors.lastname: Lee
authors.forename: Yoo Kyum
authors.initials: YK
authors.lastname: Kim
authors.forename: Sung-Woo
authors.initials: SW
authors.lastname: Cho
authors.forename: Hyun
authors.initials: H
authors.lastname: Kim
authors.forename: DongHou
authors.initials: D
authors.lastname: Kim
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins; D000072224:Bcl-2-Like Protein 11; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D000089622:Doublecortin Domain Proteins; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; D002352:Carrier Proteins; D000089622:Doublecortin Domain Proteins; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins