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

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Northern
  • DNA Primers / genetics
  • Fluorescent Antibody Technique
  • Genotype
  • HeLa Cells
  • Humans
  • Long Interspersed Nucleotide Elements / genetics*
  • Melanocytes / cytology
  • Melanocytes / metabolism*
  • Mice
  • Mice, Transgenic
  • Mutagenesis, Insertional / methods*
  • Polymerase Chain Reaction
  • Real-Time Polymerase Chain Reaction
  • Retroelements / genetics*
  • Tetracycline

Substances

  • DNA Primers
  • Retroelements
  • Tetracycline