PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells

You Tzung Chen, Kenryo Furushima, Pei Shan Hou, Amy T. Ku, Jian Min Deng, Chuan Wei Jang, Haotian Fang, Henry P. Adams, Min Liang Kuo, Hong Nerng Ho, Chung Liang Chien, Richard R. Behringer

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Permanent and reversible genetic modifications are important approaches to study gene function in different cell types. They are also important for stem cell researchers to explore and test the therapeutic potential of stem cells. The piggyBac transposon from insects is a rising nonviral system that efficiently mutagenizes and mediates gene transfer into the mammalian genome. It is also characterized by its precise excision, leaving no trace sequence behind so that the genomic integrity of the mutated cell can be restored. Here, we use an optimized piggyBac transposon system to mediate gene transfer and expression of a bifunctional fluorescent reporter in human embryonic stem (ES) cells. We provide molecular evidence for transposase-mediated piggyBac integration events and functional evidence for successful expression of a transferred fluorescent protein genes in human ES cells and their in vitro differentiated derivatives. We also demonstrate that the integrated piggyBac transposon can be removed and an undisrupted insertion site can be restored, which implies potential applications for its use in gene therapy and genetics studies.

Original languageEnglish
Pages (from-to)763-771
Number of pages9
JournalStem Cells and Development
Volume19
Issue number6
DOIs
Publication statusPublished - Jun 1 2010

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Developmental Gene Expression Regulation
Molecular Sequence Data
Transposases
Gene Transfer Techniques
DNA Transposable Elements
Cell Shape
Embryonic Stem Cells
Transgenes
Cell Differentiation
Chickens
Stem Cells
Cell Line
Genetic Therapy
Genes
Insects
Research Personnel
Genome
Gene Expression
Proteins
Human Embryonic Stem Cells

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

Cite this

Chen, Y. T., Furushima, K., Hou, P. S., Ku, A. T., Deng, J. M., Jang, C. W., ... Behringer, R. R. (2010). PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells. Stem Cells and Development, 19(6), 763-771. https://doi.org/10.1089/scd.2009.0118

PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells. / Chen, You Tzung; Furushima, Kenryo; Hou, Pei Shan; Ku, Amy T.; Deng, Jian Min; Jang, Chuan Wei; Fang, Haotian; Adams, Henry P.; Kuo, Min Liang; Ho, Hong Nerng; Chien, Chung Liang; Behringer, Richard R.

In: Stem Cells and Development, Vol. 19, No. 6, 01.06.2010, p. 763-771.

Research output: Contribution to journalArticle

Chen, YT, Furushima, K, Hou, PS, Ku, AT, Deng, JM, Jang, CW, Fang, H, Adams, HP, Kuo, ML, Ho, HN, Chien, CL & Behringer, RR 2010, 'PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells', Stem Cells and Development, vol. 19, no. 6, pp. 763-771. https://doi.org/10.1089/scd.2009.0118
Chen, You Tzung ; Furushima, Kenryo ; Hou, Pei Shan ; Ku, Amy T. ; Deng, Jian Min ; Jang, Chuan Wei ; Fang, Haotian ; Adams, Henry P. ; Kuo, Min Liang ; Ho, Hong Nerng ; Chien, Chung Liang ; Behringer, Richard R. / PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells. In: Stem Cells and Development. 2010 ; Vol. 19, No. 6. pp. 763-771.
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