title: Direct uptake of Antagomirs and efficient knockdown of miRNA in primary B and T lymphocytes.
abstract: Depending on their physiological expression level, microRNAs (miRNA) address different target genes, thus have different biological functions. To identify these, the physiological expression has to be blocked. Here, we describe the use of inhibitory cholesterol-modified oligonucleotides (Antagomirs) to inhibit miRNAs selectively in primary human and murine T and B lymphocytes. Due to their lipophilic cholesterol tag Antagomirs enter primary lymphocytes efficiently and quantitatively. We show here that at concentrations of 0.125 to 1μM, Antagomirs selectively inhibit expression of their target miRNA up to 99.5% without affecting cell viability.
authors.affiliation: Cell Biology, Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), a Leibniz institute, Berlin, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Haftmann
authors.affiliation: Cell Biology, Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), a Leibniz institute, Berlin, Germany.
authors.forename: René
authors.initials: R
authors.lastname: Riedel
authors.affiliation: Division of Molecular Immunology, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Porstner
authors.affiliation: Division of Molecular Immunology, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Wittmann
authors.affiliation: Cell Biology, Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), a Leibniz institute, Berlin, Germany.
authors.forename: Hyun-Dong
authors.initials: HD
authors.lastname: Chang
authors.affiliation: Cell Biology, Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), a Leibniz institute, Berlin, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Radbruch
authors.affiliation: Cell Biology, Deutsches Rheuma-Forschungszentrum Berlin (DRFZ), a Leibniz institute, Berlin, Germany. Electronic address: mashreghi@drfz.de.
authors.forename: Mir-Farzin
authors.initials: MF
authors.lastname: Mashreghi
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D001692:Biological Transport; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D009841:Oligonucleotides / Q000235:genetics*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology*
chemical_list: D035683:MicroRNAs; D009841:Oligonucleotides
keywords: Antagomir; Murine and human primary lymphocytes; miRNA knockdown