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  Comparative Analysis of Transposable Element Vector Systems in Human Cells.

Grabundzija, I., Irgang, M., Lajos Mátés, L. M., Belay, E., Matrai, J., Gogol-Döring, A., et al. (2010). Comparative Analysis of Transposable Element Vector Systems in Human Cells. Molecular Therapy: the Journal of the American Society of Gene Therapy, 18(6), 1200-1209. doi:10.1038/mt.2010.47.

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Genre: Zeitschriftenartikel
Alternativer Titel : Mol. Ther.

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mt201047a.pdf (beliebiger Volltext), 528KB
 
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 Urheber:
Grabundzija, Ivana, Autor
Irgang, Markus, Autor
Lajos Mátés, Lajos Mátés, Autor
Belay, Eyayu, Autor
Matrai, Janka, Autor
Gogol-Döring, Andreas, Autor
Kawakami, Koichi, Autor
Chen, Wei1, Autor           
Ruiz, Patricia, Autor
Chuah, Marinee K. L., Autor
VandenDriessche, Thierry, Autor
Izsvák, Zsuzsanna, Autor
Ivics, Zoltán, Autor
Affiliations:
1Dept. of Human Molecular Genetics (Head: Hans-Hilger Ropers), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433549              

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 Zusammenfassung: Transposon-based gene vectors have become indispensable tools in vertebrate genetics for applications ranging from insertional mutagenesis and transgenesis in model species to gene therapy in humans. The transposon toolkit is expanding, but a careful, side-by-side characterization of the diverse transposon systems has been lacking. Here we compared the Sleeping Beauty (SB), piggyBac (PB), and Tol2 transposons with respect to overall activity, overproduction inhibition (OPI), target site selection, transgene copy number as well as long-term expression in human cells. SB was the most efficient system under conditions where the availability of the transposon DNA is limiting the transposition reaction including hard-to-transfect hematopoietic stem/progenitor cells (HSCs), and the most sensitive to OPI, underpinning the need for careful optimization of the transposon components. SB and PB were about equally active, and both more efficient than Tol2, under nonrestrictive conditions. All three systems provided long-term transgene expression in human cells with minimal signs of silencing. Indeed, mapping of Tol2 insertion sites revealed significant underrepresentation within chromosomal regions with H3K27me3 histone marks typically associated with transcriptionally repressed heterochromatin. SB, Tol2, and PB constitute complementary research tools for gene transfer in mammalian cells with important implications for fundamental and translational research.

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Sprache(n): eng - English
 Datum: 2010-04-06
 Publikationsstatus: Erschienen
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Titel: Molecular Therapy : the Journal of the American Society of Gene Therapy
  Alternativer Titel : Mol. Ther.
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: -
Seiten: - Band / Heft: 18 (6) Artikelnummer: - Start- / Endseite: 1200 - 1209 Identifikator: ISSN: 1525-0016