Characterization of Engineered L1 Retrotransposition Events: The Recovery Method

David Cano; Santiago Morell; Andres J Pulgarin; Suyapa Amador; Jose L Garcia-Pérez

doi:10.1007/978-1-4939-3372-3_12

Characterization of Engineered L1 Retrotransposition Events: The Recovery Method

David Cano, Santiago Morell, Andres J Pulgarin, Suyapa Amador, Jose L Garcia-Pérez

Research output: Chapter in Book/Report/Conference proceeding › Other chapter contribution

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.

Original language	English
Title of host publication	Transposons and Retrotransposons
Publisher	Springer
Pages	165-82
Number of pages	18
Volume	1400
DOIs	https://doi.org/10.1007/978-1-4939-3372-3_12
Publication status	Published - 20 Feb 2016

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Name	Methods in Molecular Biology

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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.",

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TY - CHAP

T1 - Characterization of Engineered L1 Retrotransposition Events

T2 - The Recovery Method

AU - Cano, David

AU - Morell, Santiago

AU - Pulgarin, Andres J

AU - Amador, Suyapa

AU - Garcia-Pérez, Jose L

PY - 2016/2/20

Y1 - 2016/2/20

N2 - 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.

AB - 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.

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DO - 10.1007/978-1-4939-3372-3_12

M3 - Other chapter contribution

C2 - 26895054

VL - 1400

T3 - Methods in Molecular Biology

SP - 165

EP - 182

BT - Transposons and Retrotransposons

PB - Springer

ER -

Characterization of Engineered L1 Retrotransposition Events: The Recovery Method

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