Cationic phosphonolipids as non viral vectors for DNA transfection in hematopoietic cell lines and CD34+ cells

被引:46
作者
Floch, V
LeBolch, G
Audrezet, MP
Yaouanc, JJ
Clement, JC
desAbbayes, H
Mercier, B
Abgrall, JF
Ferec, C
机构
[1] UNIV HOSP,CTR BIOGENET,ETSBO,F-29275 BREST,FRANCE
[2] FAC SCI BREST,LAB CHIM ORGAN,UBO,URA CNRS 322,F-29275 BREST,FRANCE
[3] CHU MORVAN,SERV HEMATOL CLIN,F-29200 BREST,FRANCE
关键词
non viral vectors; gene transfer; hematopoietic cells; cationic phosphonolipids; gene therapy;
D O I
10.1006/bcmd.1997.0123
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The ability to transfer genes into a hematopoietic stem cell and to achieve regulation of their expression in lymphoid or myeloid lineages should open many new therapeutic opportunities. Besides gene transfer mediated by virus vectors like retrovirus or adenovirus, non viral systems have the theoretical advantage of being safe and easy to manage, We developed a new family of cationic lipids called phosphonolipids, synthesized 24 new molecules, and then in a first step we tested their potential to transfer genes in human hematopoietic cell lines (K562 and TF1). A LacZ plasmid under the control of a strong viral promoter was used as a reporter gene and a FAGS-Gal assay and a quantitative test CPRG assay evaluated the beta gal expression. The targeted cells were analyzed 48 hours after transfection, The present work shows that seven novel molecules display a high transfer efficiency. One of them is nine-fold more efficient than the commercially available cationic lipids. The results obtained ex vivo on CD34 cells with the FAGS-Gal assay show that at day 10 after transfection, 45 percent of cells are expressing gal.
引用
收藏
页码:69 / 87
页数:19
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