title: Cellular Localization of Engineered Human LINE-1 RNA and Proteins.
abstract: The human LINE-1 retrotransposon has the ability to mobilize into a new genomic location through an intracellular replication cycle. Immunofluorescence and in situ hybridization experiments have been developed to detect subcellular localization of retrotransposition intermediates (i.e., ORF1p, ORF2p, and L1 mRNA). Currently, these protocols are also used to validate the interaction between retrotransposition complex components and potential cellular partners involved in L1 replication. Here, we describe in details methods for the identification of LINE-1 proteins and/or RNA in cells transfected with vectors expressing engineered human LINE-1 elements.
authors.affiliation: Institute for Research on Cancer and Aging of Nice (IRCAN), CNRS (UMR 7284), INSERM (U1081), University of Nice-Sophia-Antipolis, Nice cedex 2, France.
authors.forename: Aurélien J
authors.initials: AJ
authors.lastname: Doucet
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS (UMR 5535), Montpellier Cedex 5, France.
authors.forename: Eugénia
authors.initials: E
authors.lastname: Basyuk
authors.affiliation: Institute for Regenerative Medicine and Biotherapy, INSERM (U1183), 80 Avenue Augustin Fliche, 34295, Montpellier, France. nicolas.gilbert@inserm.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Gilbert
mesh_terms: D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D005818:Genetic Engineering; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D042541:Intracellular Space / Q000378:metabolism; D020084:Long Interspersed Nucleotide Elements; D016366:Open Reading Frames; D014176:Protein Biosynthesis; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D012333:RNA, Messenger
keywords: Immunofluorescence; L1 mRNA; LINE-1 retrotransposon; MS2 fluorescent in situ hybridization; ORF1p; ORF2p