title: Characterization of Engineered L1 Retrotransposition Events: The Recovery Method.
abstract: 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.
authors.affiliation: Department of Human DNA Variability, Pfizer/University of Granada and Andalusian Regional Government Center for Genomics and Oncology (GENYO), Avda Ilustracion 114, PTS Granada, 18016, Granada, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Cano
authors.affiliation: Department of Human DNA Variability, Pfizer/University of Granada and Andalusian Regional Government Center for Genomics and Oncology (GENYO), Avda Ilustracion 114, PTS Granada, 18016, Granada, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Morell
authors.affiliation: Department of Human DNA Variability, Pfizer/University of Granada and Andalusian Regional Government Center for Genomics and Oncology (GENYO), Avda Ilustracion 114, PTS Granada, 18016, Granada, Spain.
authors.forename: Andres J
authors.initials: AJ
authors.lastname: Pulgarin
authors.affiliation: Department of Human DNA Variability, Pfizer/University of Granada and Andalusian Regional Government Center for Genomics and Oncology (GENYO), Avda Ilustracion 114, PTS Granada, 18016, Granada, Spain.
authors.forename: Suyapa
authors.initials: S
authors.lastname: Amador
authors.affiliation: Department of Human DNA Variability, Pfizer/University of Granada and Andalusian Regional Government Center for Genomics and Oncology (GENYO), Avda Ilustracion 114, PTS Granada, 18016, Granada, Spain. jlgp@genyo.es.|Genyo (Center for Genomics and Oncological Research), Pfizer/Universidad de Granada/Junta de Andalucia, PTS Granada, Spain. jlgp@genyo.es.|Institute of Genetics and Molecular Medicine (IGMM), University of Edinburgh, Edinburgh, UK. jlgp@genyo.es.
authors.forename: Jose L
authors.initials: JL
authors.lastname: Garcia-Pérez
mesh_terms: D000818:Animals; D023281:Genomics / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA
keywords: Deletion; Engineered; Insertion; LINE-1; Recovery; Retrotransposon; Target site duplication