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.
U2 - 10.1007/978-1-4939-3372-3_12
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 -