Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates

L. Mátés, Marinee K L Chuah, Eyayu Belay, Boris Jerchow, Namitha Manoj, Abel Acosta-Sanchez, Dawid P. Grzela, Andrea Schmitt, Katja Becker, Janka Matrai, Ling Ma, Ermira Samara-Kuko, Conny Gysemans, Diana Pryputniewicz, Csaba Miskey, Bradley Fletcher, Thierry Vandendriessche, Zoltán Ivics, Zsuzsanna Izsvák

Research output: Contribution to journalArticle

430 Citations (Scopus)

Abstract

The Sleeping Beauty (SB) transposon is a promising technology platform for gene transfer in vertebrates; however, its efficiency of gene insertion can be a bottleneck in primary cell types. A large-scale genetic screen in mammalian cells yielded a hyperactive transposase (SB100X) with 100-fold enhancement in efficiency when compared to the first-generation transposase. SB100X supported 35-50% stable gene transfer in human CD34 + cells enriched in hematopoietic stem or progenitor cells. Transplantation of gene-marked CD34 + cells in immunodeficient mice resulted in long-term engraftment and hematopoietic reconstitution. In addition, SB100X supported sustained (1 year) expression of physiological levels of factor IX upon transposition in the mouse liver in vivo. Finally, SB100X reproducibly resulted in 45% stable transgenesis frequencies by pronuclear microinjection into mouse zygotes. The newly developed transposase yields unprecedented stable gene transfer efficiencies following nonviral gene delivery that compare favorably to stable transduction efficiencies with integrating viral vectors and is expected to facilitate widespread applications in functional genomics and gene therapy.

Original languageEnglish
Pages (from-to)753-761
Number of pages9
JournalNature Genetics
Volume41
Issue number6
DOIs
Publication statusPublished - Jun 2009

Fingerprint

Transposases
Beauty
Molecular Evolution
Vertebrates
Genes
Hematopoietic Stem Cells
Gene Transfer Techniques
Factor IX
Zygote
Insertional Mutagenesis
Microinjections
Genomics
Genetic Therapy
Transplantation
Technology
Liver

ASJC Scopus subject areas

  • Genetics
  • Medicine(all)

Cite this

Mátés, L., Chuah, M. K. L., Belay, E., Jerchow, B., Manoj, N., Acosta-Sanchez, A., ... Izsvák, Z. (2009). Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nature Genetics, 41(6), 753-761. https://doi.org/10.1038/ng.343

Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. / Mátés, L.; Chuah, Marinee K L; Belay, Eyayu; Jerchow, Boris; Manoj, Namitha; Acosta-Sanchez, Abel; Grzela, Dawid P.; Schmitt, Andrea; Becker, Katja; Matrai, Janka; Ma, Ling; Samara-Kuko, Ermira; Gysemans, Conny; Pryputniewicz, Diana; Miskey, Csaba; Fletcher, Bradley; Vandendriessche, Thierry; Ivics, Zoltán; Izsvák, Zsuzsanna.

In: Nature Genetics, Vol. 41, No. 6, 06.2009, p. 753-761.

Research output: Contribution to journalArticle

Mátés, L, Chuah, MKL, Belay, E, Jerchow, B, Manoj, N, Acosta-Sanchez, A, Grzela, DP, Schmitt, A, Becker, K, Matrai, J, Ma, L, Samara-Kuko, E, Gysemans, C, Pryputniewicz, D, Miskey, C, Fletcher, B, Vandendriessche, T, Ivics, Z & Izsvák, Z 2009, 'Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates', Nature Genetics, vol. 41, no. 6, pp. 753-761. https://doi.org/10.1038/ng.343
Mátés, L. ; Chuah, Marinee K L ; Belay, Eyayu ; Jerchow, Boris ; Manoj, Namitha ; Acosta-Sanchez, Abel ; Grzela, Dawid P. ; Schmitt, Andrea ; Becker, Katja ; Matrai, Janka ; Ma, Ling ; Samara-Kuko, Ermira ; Gysemans, Conny ; Pryputniewicz, Diana ; Miskey, Csaba ; Fletcher, Bradley ; Vandendriessche, Thierry ; Ivics, Zoltán ; Izsvák, Zsuzsanna. / Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. In: Nature Genetics. 2009 ; Vol. 41, No. 6. pp. 753-761.
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