Characterization of Engineered L1 Retrotransposition Events: The Recovery Method

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1400; p. 165
• Main Authors: Cano, David, Morell, Santiago, Pulgarin, Andres J, Amador, Suyapa, Garcia-Pérez, Jose L
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Animals; Genomics - methods; HeLa Cells; Humans; Long Interspersed Nucleotide Elements; Open Reading Frames; Sequence Analysis, DNA; Engineered; LINE-1; Retrotransposon; Target site duplication; Deletion; Insertion; Recovery
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-3372-3_12

Long Interspersed Element class 1 retrotransposons (LINE-1 or L1) are abundant Transposable Elements in mammalian genomes and their mobility continues to impact the human genome. The development of engineered retrotransposition assays has been instrumental to understand how these elements are regulated and to identify domains involved in the process of retrotransposition. Additionally, the modification of a retrotransposition indicator cassette has allowed developing straightforward approaches to characterize the site of new L1 insertions in cultured cells. In this chapter, we describe a method termed "L1-recovery" that has been used to characterize the site of insertion on engineered L1 retrotransposition events in cultured mammalian cells. Notably, the recovery assay is based on a genetic strategy and avoids the use of PCR and thus reduces to a minimum the appearance of false positives/artifacts.