TY - JOUR
T1 - High frequency retrotransposition in cultured mammalian cells
AU - Moran, John V.
AU - Holmes, Susan Elizabeth
AU - Naas, Thierry P.
AU - DeBerardinis, Ralph J.
AU - Boeke, Jef D.
AU - Kazazian, Haig H.
N1 - Funding Information:
Correspondence should be addressed to H. H. K. We thank Drs. A. Scott, B. Dombroski, M. Singer, G. Swergold, R. Kennett, V. Lauermann, Q. Feng, and D. Sassaman for helpful discussions, Dr. D. Mager for providing the mneoI cassette prior to publication, Dr. M. Meisler for discussing results prior to publication, Dr. V. Bedian and the University of Pennsylvania DNA sequencing core for all their help, and Drs. T. Kadesch and P. Perlman for critical reading of the manuscript. This work was supported by National Institutes of Health grants to H. H. K. and J. D. B.; J. V. M. was supported by a Damon Runyon–Walter Winchell Cancer Research Fund Fellowship (DRG 1332); and T. P. N. was supported by a Long-Term European Molecular Biology Organization fellowship.
PY - 1996/11/29
Y1 - 1996/11/29
N2 - We previously isolated two human L1 elements (L1.2 and LRE2) as the progenitors of disease-producing insertions. Here, we show these elements can actively retrotranspose in cultured mammalian cells. When stably expressed from an episome in HeLa cells, both elements retrotransposed into a variety of chromosomal locations at a high frequency. The retrotransposed products resembled endogenous L1 insertions, since they were variably 5' truncated, ended in poly(A) tracts, and were flanked by target-site duplications or short deletions. Point mutations in conserved domains of the L1.2-encoded proteins reduced retrotransposition by 100- to 1000-fold. Remarkably, L1.2 also retrotransposed in a mouse cell line, suggesting a potential role for L1-based vectors in random insertional mutagenesis.
AB - We previously isolated two human L1 elements (L1.2 and LRE2) as the progenitors of disease-producing insertions. Here, we show these elements can actively retrotranspose in cultured mammalian cells. When stably expressed from an episome in HeLa cells, both elements retrotransposed into a variety of chromosomal locations at a high frequency. The retrotransposed products resembled endogenous L1 insertions, since they were variably 5' truncated, ended in poly(A) tracts, and were flanked by target-site duplications or short deletions. Point mutations in conserved domains of the L1.2-encoded proteins reduced retrotransposition by 100- to 1000-fold. Remarkably, L1.2 also retrotransposed in a mouse cell line, suggesting a potential role for L1-based vectors in random insertional mutagenesis.
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U2 - 10.1016/S0092-8674(00)81998-4
DO - 10.1016/S0092-8674(00)81998-4
M3 - Article
C2 - 8945518
AN - SCOPUS:0030606320
SN - 0092-8674
VL - 87
SP - 917
EP - 927
JO - Cell
JF - Cell
IS - 5
ER -