title: Mitochondrial targeting of recombinant RNA.
abstract: Mitochondrial import of small noncoding RNA is found in a large variety of species. In mammalian cells, this pathway can be used for therapeutic purpose, to restore the mitochondrial functions affected by pathogenic mutations. Recently, we developed mitochondrial RNA vectors able to address therapeutic oligoribonucleotides into human mitochondria. Here we provide the protocol for transfection of cultured human cells with small recombinant RNA molecules and describe two approaches useful to demonstrate their import into mitochondria: (1) isolation of RNA from purified mitochondria and quantitative hybridization analysis and (2) confocal microscopy of cells transfected with fluorescently labeled RNA. These protocols can be used in combination with overexpression or downregulation of protein import factors to detect and to evaluate their influence on the mitochondrial import of various RNAs.
authors: UMR 7156 Genetique Moleculaire, Genomique, Microbiologie (GMGM), University of Strasbourg-CNRS, 21 Rue René Descartes, 67084, Strasbourg, France.
authors: Ilya
authors: I
authors: Dovydenko
authors: Anne-Marie
authors: AM
authors: Heckel
authors: Yann
authors: Y
authors: Tonin
authors: Ali
authors: A
authors: Gowher
authors: Alya
authors: A
authors: Venyaminova
authors: Ivan
authors: I
authors: Tarassov
authors: Nina
authors: N
authors: Entelis
mesh_terms: D002478:Cells, Cultured; D015870:Gene Expression; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism*; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D034443:RNA Transport; D000077278:RNA, Mitochondrial; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
chemical_list: D000077278:RNA, Mitochondrial; D034741:RNA, Small Interfering; D012313:RNA