High Frequency Retrotransposition in Cultured Mammalian Cells

作者：John V. Moran, Susan E. Holmes, Thierry Naas, Ralph J. DeBerardinis, Jef D. Boeke, Haig H. Kazazian · 发表于：Cell · 年份：1996 · DOI：10.1016/s0092-8674(00)81998-4 · 被引用次数：1049 · 研究领域：Chromosomal and Genetic Variations、Genomics and Chromatin Dynamics、CRISPR and Genetic Engineering

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.