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T Cell‐specific expression from Mo‐MLV retroviral vectors containing a CD4 mini‐promoter/enhancer
Author(s) -
ZhaoEmonet Jing Chao,
Marodon Gilles,
PiocheDurieu Catherine,
Cosset François Loic,
Klatzmann David
Publication year - 2000
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/1521-2254(200011/12)2:6<416::aid-jgm142>3.0.co;2-y
Subject(s) - enhancer , murine leukemia virus , viral vector , biology , green fluorescent protein , microbiology and biotechnology , expression cassette , vector (molecular biology) , transfection , flow cytometry , gene , titer , genetic enhancement , virology , virus , gene expression , genetics , recombinant dna
Background Gene therapy of various immunological disorders will greatly benefit from improved retroviral vectors (RVs) with T cell specificity. Such vectors can be designed by placing a gene of therapeutic interest under the control of tissue‐specific transcriptional elements. However, low titers and loss of specificity are frequently encountered with tissue‐specific vectors. The aim of the present study was to develop a T cell‐specific RV. Methods We constructed a series of Moloney murine leukemia virus (Mo‐MLV)‐based RVs expressing enhanced green fluorescent protein (EGFP) under the control of a mini‐promoter/enhancer cassette derived from the CD4 gene (CD4pmE) and tested them in cell lines and peripheral blood lymphocytes. Expression of EGFP was monitored by fluorescence microscopy and analyzed by flow cytometry. Results The CD4pmE cassette was inserted between the viral long terminal repeats (LTRs) in self‐inactivating vectors (SIN vectors) or was substituted to the 3′ U3 viral promoter/enhancer (hybrid vectors). High vector titers but poor specific expression of EGFP were achieved when CD4pmE was inserted in sense orientation in SIN vectors. Low titers but high specificity were observed when the CD4pmE cassette was in anti‐sense orientation. In contrast, high titers and good T cell specificity were obtained with hybrid vectors. Conclusion An efficient T cell‐specific retroviral vector was obtained. Copyright © 2000 John Wiley & Sons, Ltd.