title: LINE-1 Cultured Cell Retrotransposition Assay.
abstract: The Long INterspersed Element-1 (LINE-1 or L1) retrotransposition assay has facilitated the discovery and characterization of active (i.e., retrotransposition-competent) LINE-1 sequences from mammalian genomes. In this assay, an engineered LINE-1 containing a retrotransposition reporter cassette is transiently transfected into a cultured cell line. Expression of the reporter cassette, which occurs only after a successful round of retrotransposition, allows the detection and quantification of the LINE-1 retrotransposition efficiency. This assay has yielded insight into the mechanism of LINE-1 retrotransposition. It also has provided a greater understanding of how the cell regulates LINE-1 retrotransposition and how LINE-1 retrotransposition impacts the structure of mammalian genomes. Below, we provide a brief introduction to LINE-1 biology and then detail how the LINE-1 retrotransposition assay is performed in cultured mammalian cells.
authors.affiliation: Department of Human Genetics, University of Michigan Medical School, 1241 E. Catherine St, Ann Arbor, MI, 48109, USA. chongh@umich.edu.
authors.forename: Huira C
authors.initials: HC
authors.lastname: Kopera
authors.affiliation: Department of Human Genetics, University of Michigan Medical School, 1241 E. Catherine St, Ann Arbor, MI, 48109, USA.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Larson
authors.affiliation: Cellular and Molecular Biology Program, University of Michigan Medical School, 1241 E. Catherine St, Ann Arbor, MI, 48109, USA.
authors.forename: John B
authors.initials: JB
authors.lastname: Moldovan
authors.affiliation: Department of Human Genetics, University of Michigan Medical School, 1241 E. Catherine St, Ann Arbor, MI, 48109, USA.
authors.forename: Sandra R
authors.initials: SR
authors.lastname: Richardson
authors.affiliation: Howard Hughes Medical Institute, Chevy Chase, MD, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Human Genetics, University of Michigan Medical School, 1241 E. Catherine St, Ann Arbor, MI, 48109, USA. moranj@umich.edu.|Cellular and Molecular Biology Program, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. moranj@umich.edu.|Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. moranj@umich.edu.|Howard Hughes Medical Institute, Chevy Chase, MD, USA. moranj@umich.edu.
authors.forename: John V
authors.initials: JV
authors.lastname: Moran
mesh_terms: D020087:Alu Elements; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D018626:Retroelements; D014162:Transfection
chemical_list: D018626:Retroelements
keywords: Alu; LINE-1; Mammalian cultured cells; Retrotransposition assay; trans-complementation assay