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

Andrew Wilber, Joel L. Frandsen, Jennifer L. Geurts, David A. Largaespada, Perry B. Hackett, R. Scott McIvor

Research output: Contribution to journalArticlepeer-review

95 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)625-630
Number of pages6
JournalMolecular Therapy
Volume13
Issue number3
DOIs
StatePublished - Mar 2006

Bibliographical note

Funding Information:
We thank Aron Geurts, Corey Carlson, and the Biological Imaging and Processing Laboratory at the University of Minnesota for technical assistance. This work was supported by a grant from the Arnold and Mabel Beckman Foundation (R.S.M) and Training Grant T32 GM08347 from NIGMS (A.W.). A.W. is the recipient of a University of Minnesota Doctoral Dissertation Fellowship.

Keywords

  • Bioluminescent imaging
  • Liver
  • Non-viral integration
  • RNA delivery
  • Sleeping Beauty

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