Characterization of Engineered L1 Retrotransposition Events: The Recovery Method

Autor:	Andres J Pulgarin, David Cano, Santiago Morell, Suyapa Amador, Jose L. Garcia-Perez
Rok vydání:	2016
Předmět:	0301 basic medicine Long interspersed nuclear element Transposable element 03 medical and health sciences Open reading frame 030104 developmental biology Recovery method Retrotransposon Human genome Genomics Computational biology Biology Genome
Zdroj:	Methods in Molecular Biology ISBN: 9781493933709 Transposons and Retrotransposons
DOI:	10.1007/978-1-4939-3372-3_12
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::40674dc30e6c16d71c84ee408d1dc419 https://doi.org/10.1007/978-1-4939-3372-3_12 Zobrazit plný text záznamu