RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues

Mol Ther. 2006 Mar;13(3):625-30. doi: 10.1016/j.ymthe.2005.10.014. Epub 2005 Dec 20.


Sleeping Beauty (SB) is a DNA transposon capable of mediating gene insertion and long-term expression in vertebrate cells when co-delivered with a source of transposase. In all previous reports of SB-mediated gene insertion in somatic cells, the transposase component has been provided by expression of a co-delivered DNA molecule that has the potential for integration into the host cell genome. Integration and continued expression of a gene encoding SB transposase could be problematic if it led to transposon re-mobilization and reintegration. We addressed this potential problem by supplying the transposase-encoding molecule in the form of mRNA. We show that transposase-encoding mRNA can effectively mediate transposition in vitro in HT1080 cells and in vivo in mouse liver following co-delivery with a recoverable transposon or with a luciferase transposon. We conclude that in vitro-transcribed mRNA can be used as an effective source of transposase for SB-mediated transposition in mammalian cells and tissues.

Publication types

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

MeSH terms

  • Animals
  • Cell Line, Tumor
  • DNA Transposable Elements*
  • Gene Expression Regulation*
  • Gene Transfer Techniques*
  • Humans
  • Liver / enzymology
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Organ Specificity / genetics
  • RNA, Messenger / administration & dosage
  • RNA, Messenger / genetics*
  • Recombination, Genetic
  • Transposases / genetics*


  • DNA Transposable Elements
  • RNA, Messenger
  • Transposases
  • sleeping beauty transposase, human