Messenger RNA as a source of transposase for Sleeping Beauty transposon-mediated correction of hereditary tyrosinemia type I

Andrew Wilber, Kirk J. Wangensteen, Yixin Chen, Lijuan Zhuo, Joel L. Frandsen, Jason B. Bell, Zongyu J. Chen, Stephen C Ekker, R. Scott McIvor, Xin Wang

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

The Sleeping Beauty (SB) transposon system mediates chromosomal integration and stable gene expression when an engineered SB transposon is delivered along with transposase. One concern in the therapeutic application of the SB system is that persistent expression of transposase could result in transposon instability and genotoxicity. Here, we tested the use of transposase-encoding RNA plus transposon DNA for correction of murine fumarylacetoacetate hydrolase (FAH) deficiency. A bi-functional transposon containing both mouse FAH and firefly luciferase sequences was used to track the growth of genetically corrected liver tissue by in vivo bioluminescence imaging after delivery of DNA or RNA as a source of transposase. Supplying SB transposase in the form of RNA resulted in selective repopulation of corrected hepatocytes with stable expression of FAH and luciferase. Plasma succinylacetone and amino acid levels were normalized, suggesting normal liver metabolism of catabolized protein products. Secondary FAH-deficient animals transplanted with hepatocytes (250,000) isolated from primary treated animals survived 2-(2-nitro-4-trifluoro-methylbenzoyl)-1,3-cyclohexanedione (NTBC) withdrawal, gained weight consistently, and demonstrated stable expression of luciferase. We conclude that transposase-encoding messenger RNA (mRNA) can be used to mediate stable non-viral gene therapy, resulting in complete phenotypic correction, and is thus an effective source of recombinase activity for use in human gene therapy.

Original languageEnglish (US)
Pages (from-to)1280-1287
Number of pages8
JournalMolecular Therapy
Volume15
Issue number7
DOIs
StatePublished - Jul 2007
Externally publishedYes

Fingerprint

Tyrosinemias
Transposases
Beauty
Messenger RNA
RNA
Luciferases
Genetic Therapy
Hepatocytes
Firefly Luciferases
Recombinases
DNA Transposable Elements
Liver
Gene Expression
Amino Acids
Weights and Measures
fumarylacetoacetase
DNA
Growth

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Wilber, A., Wangensteen, K. J., Chen, Y., Zhuo, L., Frandsen, J. L., Bell, J. B., ... Wang, X. (2007). Messenger RNA as a source of transposase for Sleeping Beauty transposon-mediated correction of hereditary tyrosinemia type I. Molecular Therapy, 15(7), 1280-1287. https://doi.org/10.1038/sj.mt.6300160

Messenger RNA as a source of transposase for Sleeping Beauty transposon-mediated correction of hereditary tyrosinemia type I. / Wilber, Andrew; Wangensteen, Kirk J.; Chen, Yixin; Zhuo, Lijuan; Frandsen, Joel L.; Bell, Jason B.; Chen, Zongyu J.; Ekker, Stephen C; McIvor, R. Scott; Wang, Xin.

In: Molecular Therapy, Vol. 15, No. 7, 07.2007, p. 1280-1287.

Research output: Contribution to journalArticle

Wilber, A, Wangensteen, KJ, Chen, Y, Zhuo, L, Frandsen, JL, Bell, JB, Chen, ZJ, Ekker, SC, McIvor, RS & Wang, X 2007, 'Messenger RNA as a source of transposase for Sleeping Beauty transposon-mediated correction of hereditary tyrosinemia type I', Molecular Therapy, vol. 15, no. 7, pp. 1280-1287. https://doi.org/10.1038/sj.mt.6300160
Wilber, Andrew ; Wangensteen, Kirk J. ; Chen, Yixin ; Zhuo, Lijuan ; Frandsen, Joel L. ; Bell, Jason B. ; Chen, Zongyu J. ; Ekker, Stephen C ; McIvor, R. Scott ; Wang, Xin. / Messenger RNA as a source of transposase for Sleeping Beauty transposon-mediated correction of hereditary tyrosinemia type I. In: Molecular Therapy. 2007 ; Vol. 15, No. 7. pp. 1280-1287.
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