title: Lentiviral overexpression of miRNAs.
abstract: Deregulation of microRNAs (miRNAs) has been attributed to almost any human disease analyzed to date. This calls for models and experimental strategies for functional analyses of miRNAs enabling miRNA overexpression or suppression in target cell and/or tissues. Lentiviral vector (LV)-based technologies allow the long-term overexpression of miRNAs in nearly all cell types in an easy and rapid manner and are therefore popular tools for this application. In this chapter we describe the cloning of LV miRNA expression vectors as well as the production of virus particles for target cell infection and stable expression of miRNAs.
authors.affiliation: Labor für Molekulare Gastroenterologische Onkologie (MGO), Zentrum für Klinische Forschung (ZKF), Ruhr Universität Bochum, Bochum, Germany.
authors.forename: Hannah
authors.initials: H
authors.lastname: Zöllner
authors.forename: Stephan A
authors.initials: SA
authors.lastname: Hahn
authors.forename: Abdelouahid
authors.initials: A
authors.lastname: Maghnouj
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic
chemical_list: D017931:DNA Primers; D035683:MicroRNAs