LINE-1 Cultured Cell Retrotransposition Assay

Methods Mol Biol. 2016:1400:139-56. doi: 10.1007/978-1-4939-3372-3_10.

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.

Keywords: Alu; LINE-1; Mammalian cultured cells; Retrotransposition assay; trans-complementation assay.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alu Elements
  • Gene Expression
  • HeLa Cells
  • Humans
  • Long Interspersed Nucleotide Elements*
  • Retroelements
  • Transfection

Substances

  • Retroelements