| Chapter title |
LINE-1 Cultured Cell Retrotransposition Assay.
|
|---|---|
| Chapter number | 10 |
| Book title |
Transposons and Retrotransposons
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3372-3_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3370-9, 978-1-4939-3372-3
|
| Authors |
Huira C. Kopera Ph.D., Peter A. Larson, John B. Moldovan, Sandra R. Richardson, Ying Liu, John V. Moran Ph.D., Huira C. Kopera, John V. Moran |
| Editors |
Jose L. Garcia-Pérez |
| 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. |
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