|
|
Blood, 15 January 2006, Vol. 107, No. 2, pp. 483-491.
Prepublished online as a Blood First Edition Paper on September 27, 2005; DOI 10.1182/blood-2005-05-2133.
Previous Article | Next Article 
Submitted May 26, 2005
Accepted September 10, 2005
Stable gene transfer and expression in human primary T-cells by the Sleeping Beauty transposon system
Xin Huang, Andrew C Wilber, Lei Bao, Dong Tuong, Jakub Tolar, Paul J Orchard, Bruce L Levine, Carl H June, R S McIvor, Bruce R Blazar, and Xianzheng Zhou*
Division of Blood and Marrow Transplantation, Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA; The Cancer Center, University of Minnesota, Minneapolis, MN, USA
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA; The Cancer Center, University of Minnesota, Minneapolis, MN, USA
Department of Pathology and Laboratory Medicine, the Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, PA, USA
* Corresponding author; email: zhoux058{at}umn.edu.
The Sleeping Beauty (SB) transposon system is a non-viral DNA delivery system in which a transposase directs integration of an SB transposon into TA-dinucleotide sites in the genome. To determine whether the SB transposon system can mediate stable gene expression in human T-cells, primary peripheral blood lymphocytes (PBLs) were nucleofected with SB vectors carrying a DsRed reporter gene. Plasmids containing the SB transposase on the same (cis) or separate molecule (trans) as the SB transposon mediated long-term and stable reporter gene expression in human primary T-cells. Sequencing of transposon:chromosome junctions confirmed that stable gene expression was due to SB-mediated transposition. In other studies, PBLs were successfully transfected using the SB transposon system and shown to stably express a fusion protein consisting of (i) a surface receptor useful for positive T-cell selection and (ii) a "suicide" gene useful for elimination of transfected T-cells after chemotherapy. This study is the first report demonstrating that the SB transposon system can mediate stable gene transfer in human primary PBLs, which may be advantageous for T-cell based gene therapies.

CiteULike Connotea Del.icio.us Digg Reddit Technorati What's this?
Related Article in Blood Online:
-
Sleeping Beauty awakens!
- Bruce A. Bunnell
Blood 2006 107: 416-417.
[Full Text]
[PDF]
This article has been cited by other articles:

|
 |

|
 |
 
X. Xue, X. Huang, S. E. Nodland, L. Mates, L. Ma, Z. Izsvak, Z. Ivics, T. W. LeBien, R. S. McIvor, J. E. Wagner, et al.
Stable gene transfer and expression in cord blood-derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system
Blood,
August 13, 2009;
114(7):
1319 - 1330.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
H. Singh, P. R. Manuri, S. Olivares, N. Dara, M. J. Dawson, H. Huls, P. B. Hackett, D. B. Kohn, E. J. Shpall, R. E. Champlin, et al.
Redirecting Specificity of T-Cell Populations For CD19 Using the Sleeping Beauty System
Cancer Res.,
April 15, 2008;
68(8):
2961 - 2971.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. M. Doubrovin, E. S. Doubrovina, P. Zanzonico, M. Sadelain, S. M. Larson, and R. J. O'Reilly
In vivo Imaging and Quantitation of Adoptively Transferred Human Antigen-Specific T Cells Transduced to Express a Human Norepinephrine Transporter Gene
Cancer Res.,
December 15, 2007;
67(24):
11959 - 11969.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
T. Zhang, A. Barber, and C. L. Sentman
Generation of Antitumor Responses by Genetic Modification of Primary Human T Cells with a Chimeric NKG2D Receptor
Cancer Res.,
June 1, 2006;
66(11):
5927 - 5933.
[Abstract]
[Full Text]
[PDF]
|
 |
|
|
|