High Frequency Retrotransposition in Cultured Mammalian Cells
Two human L1 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.
Proposed Use (Set) 1) Insertional mutagenesis in mammalian cells 2) Gene delivery in human cells i.e., for gene therapy
Boeke, Jef D.
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