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Proc Natl Acad Sci U S A. 2013 Jul 16;110(29):E2706-13. doi: 10.1073/pnas.1302504110. Epub 2013 Jul 1.

Controlled insertional mutagenesis using a LINE-1 (ORFeus) gene-trap mouse model.

Author information

1
Department of Molecular Biology and Genetics, and High Throughput Biology Center, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. kathryn.odonnell@UTSouthwestern.edu

Abstract

A codon-optimized mouse LINE-1 element, ORFeus, exhibits dramatically higher retrotransposition frequencies compared with its native long interspersed element 1 counterpart. To establish a retrotransposon-mediated mouse model with regulatable and potent mutagenic capabilities, we generated a tetracycline (tet)-regulated ORFeus element harboring a gene-trap cassette. Here, we show that mice expressing tet-ORFeus broadly exhibit robust retrotransposition in somatic tissues when treated with doxycycline. Consistent with a significant mutagenic burden, we observed a reduced number of double transgenic animals when treated with high-level doxycycline during embryogenesis. Transgene induction in skin resulted in a white spotting phenotype due to somatic ORFeus-mediated mutations that likely disrupt melanocyte development. The data suggest a high level of transposition in melanocyte precursors and consequent mutation of genes important for melanoblast proliferation, differentiation, or migration. These findings reveal the utility of a retrotransposon-based mutagenesis system as an alternative to existing DNA transposon systems. Moreover, breeding these mice to different tet-transactivator/reversible tet-transactivator lines supports broad functionality of tet-ORFeus because of the potential for dose-dependent, tissue-specific, and temporal-specific mutagenesis.

KEYWORDS:

L1 retrotransposon; mus musculus; tet-promoter; white-spotted phenotype

PMID:
23818630
PMCID:
PMC3718180
DOI:
10.1073/pnas.1302504110
[Indexed for MEDLINE]
Free PMC Article
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