Gene transfer into genomes of human cells by the Sleeping Beauty transposon system

Aron M. Geurts, Ying Yang, Karl J. Clark, Geyi Liu, Zongbin Cui, Adam J. Dupuy, Jason B. Bell, David A. Largaespada, Perry B. Hackett

Research output: Contribution to journalArticlepeer-review

303 Scopus citations


The Sleeping Beauty (SB) transposon system, derived from teleost fish sequences, is extremely effective at delivering DNA to vertebrate genomes, including those of humans. We have examined several parameters of the SB system to improve it as a potential, nonviral vector for gene therapy. Our investigation centered on three features: the carrying capacity of the transposon for efficient integration into chromosomes of HeLa cells, the effects of overexpression of the SB transposase gene on transposition rates, and improvements in the activity of SB transposase to increase insertion rates of transgenes into cellular chromosomes. We found that SB transposons of about 6 kb retained 50% of the maximal efficiency of transposition, which is sufficient to deliver 70-80% of identified human cDNAs with appropriate transcriptional regulatory sequences. Overexpression inhibition studies revealed that there are optimal ratios of SB transposase to transposon for maximal rates of transposition, suggesting that conditions of delivery of the two-part transposon system are important for the best gene-transfer efficiencies. We further refined the SB transposase to incorporate several amino acid substitutions, the result of which led to an improved transposase called SB11. With SB11 we are able to achieve transposition rates that are about 100-fold above those achieved with plasmids that insert into chromosomes by random recombination. With the recently described improvements to the transposon itself, the SB system appears to be a potential gene-transfer tool for human gene therapy.

Original languageEnglish (US)
Pages (from-to)108-117
Number of pages10
JournalMolecular Therapy
Issue number1
StatePublished - Jul 1 2003

Bibliographical note

Funding Information:
We thank Stephen Ekker, Scott McIvor, Elena Aronovich, Betsy Kren, and members of the Beckman Center for Transposon Research for helpful analyses and discussions. This work was supported by grants from the Arnold and Mabel Beckman Foundation and the NIH (NCRR R01-066525-07 and NICHD P01-HD32652).


  • Gene therapy
  • Inverted terminal repeats
  • Site-specific mutagenesis
  • Transposase


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