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Gewählte Publikation:

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Publikationstyp: Zeitschriftenaufsatz
Dokumenttyp: Originalarbeit

Jahr: 2001

AutorInnen: Indraccolo, S; Minuzzo, S; Roccaforte, F; Zamarchi, R; Habeler, W; Stievano, L; Tosello, V; Klein, D; Günzburg, WH; Basso, G; Chieco-Bianchi, L; Amadori, A

Titel: Effects of CD2 locus control region sequences on gene expression by retroviral and lentiviral vectors.

Quelle: Blood. 2001; 98(13):3607-3617



Autor/innen der Vetmeduni Vienna:

Günzburg Walter
Klein Dieter

Beteiligte Vetmed-Organisationseinheiten
Institut für Virologie


Zugehörige(s) Projekt(e): Genomregulation und Gentherapie in Human- und Veterinärmedizin


Abstract:
Locus control region (LCR) sequences are involved in the establishment of open chromosomal domains. To evaluate the possibility of exploiting the human CD2 LCR to regulate gene expression by Moloney murine leukemia virus (Mo-MLV)-based retroviral vectors in T cells, it was included in vectors carrying the enhanced green fluorescence protein (EGFP) reporter gene; then transduction in vitro of lymphoid and nonlymphoid cell lines was performed. Deletion of the viral enhancer in the Mo-MLV long terminal repeat was necessary to detect LCR activity in the context of these retroviral vectors. It was found that a full-length (2.1 kb), but not a truncated (1.0 kb), CD2 LCR retained the ability to modulate reporter gene expression by Mo-MLV-derived retroviral vectors, leading to a homogeneous, unimodal pattern of EGFP expression that remained unmodified in culture over time, specifically in T-cell lines; on the other hand, viral titer was strongly reduced compared with vectors not carrying the LCR. Lentiviral vectors containing the CD2 LCR could be generated at higher titers and were used to analyze its effects on gene expression in primary T cells. Subcutaneous implantation of genetically modified cells in immunodeficient mice showed that retroviral vectors carrying the CD2 LCR conferred an advantage in terms of transgene expression in vivo, compared with the parental vector, by preventing the down-modulation of EGFP expression. These findings suggest a potential application of this LCR to increase gene expression by retroviral and lentiviral vectors in T lymphocytes.

Keywords Pubmed: 3T3 Cells
Animals
Antigens, CD2/genetics*
Blotting, Southern
Cell Line
Gene Expression*
Genetic Vectors*
Green Fluorescent Proteins
Humans
Kidney
Lentivirus/genetics*
Locus Control Region*
Luminescent Proteins/genetics
Mice
Moloney murine leukemia virus/genetics
Retroviridae/genetics*
T-Lymphocytes/metabolism
Transfection


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