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.
CITATION STYLE
Kopera, H. C., Larson, P. A., Moldovan, J. B., Richardson, S. R., Liu, Y., & Moran, J. V. (2016). Line-1 cultured cell retrotransposition assay. In Methods in Molecular Biology (Vol. 1400, pp. 139–156). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3372-3_10
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