Molecular Genetics and Genomics

, 282:531

Physical properties of DNA components affecting the transposition efficiency of the mariner Mos1 element

  • Sophie Casteret
  • Najat Chbab
  • Jeanne Cambefort
  • Corinne Augé-Gouillou
  • Yves Bigot
  • Florence Rouleux-Bonnin
Original Paper

DOI: 10.1007/s00438-009-0484-0

Cite this article as:
Casteret, S., Chbab, N., Cambefort, J. et al. Mol Genet Genomics (2009) 282: 531. doi:10.1007/s00438-009-0484-0
  • 136 Downloads

Abstract

Previous studies have shown that the transposase and the inverted terminal repeat (ITR) of the Mos1mariner elements are suboptimal for transposition; and that hyperactive transposases and transposon with more efficient ITR configurations can be obtained by rational molecular engineering. In an attempt to determine the extent to which this element is suboptimal for transposition, we investigate here the impact of the three main DNA components on its transposition efficiency in bacteria and in vitro. We found that combinations of natural and synthetic ITRs obtained by systematic evolution of ligands by exponential enrichment did increase the transposition rate. We observed that when untranslated terminal regions were associated with their respective natural ITRs, they acted as transposition enhancers, probably via the early transposition steps. Finally, we demonstrated that the integrity of the Mos1 inner region was essential for transposition. These findings allowed us to propose prototypes of optimized Mos1 vectors, and to define the best sequence features of their associated marker cassettes. These vector prototypes were assayed in HeLa cells, in which Mos1 vectors had so far been found to be inactive. The results obtained revealed that using these prototypes does not circumvent this problem. However, such vectors can be expected to provide new tools for the use in genome engineering in systems such as Caenorhabditis elegans in which Mos1 is very active.

Keywords

DNA curvature DNA bendability DNA binding Transposon ITm elements 

Abbreviations

HTH

Helix-turn-helix

ITR

Inverted terminal repeat

ORF

Open reading frame

SELEX

Systematic evolution of ligands by exponential enrichment

TE

Transposable element

UTR

Untranslated terminal region

Supplementary material

438_2009_484_MOESM1_ESM.doc (62 kb)
Supplementary material 1 (DOC 62 kb)

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Sophie Casteret
    • 1
    • 2
  • Najat Chbab
    • 1
    • 2
    • 4
  • Jeanne Cambefort
    • 1
    • 2
  • Corinne Augé-Gouillou
    • 1
    • 2
  • Yves Bigot
    • 1
    • 2
    • 3
  • Florence Rouleux-Bonnin
    • 1
    • 2
  1. 1.Université François Rabelais de Tours, GICCUFR des Sciences and TechniquesToursFrance
  2. 2.GICC, UMR CNRS 6239, Université François RabelaisUFR des Sciences and TechniquesToursFrance
  3. 3.CHRU de ToursUFR des Sciences and TechniquesToursFrance
  4. 4.Institut für VirologieBerlinGermany

Personalised recommendations