| Abstract: |
Fetuses with homozygous α-thalassemia usually die at the third trimester of pregnancy or soon after birth. Hence, the disease could potentially be a target for fetal gene therapy. We have previously established a mouse model of α-thalassemia. These mice mimic the human α-thalassemic conditions and can be used as preclinical models for fetal gene therapy. We tested a lentiviral vector containing the HS 2, 3, and 4 of the β-LCR, a central polypurine tract element, and the β-globin gene promoter directing either the EGFP or the human α-globin gene. We showed that the GFP expression was erythroid-specific and detected in BFU-E colonies and the erythroid progenies of CFU-GEMM. For in utero gene delivery, we did yolk sac vessel injection at midgestation of mouse embryos. The recipient mice were analyzed after birth for human α-globin gene expression. In the newborn, human α-globin gene expression was detected in the liver, spleen, and peripheral blood. The human α-globin gene expression was at the peak at 3-4 months, when it reached 20% in some recipients. However, the expression declined at 7 months. Colony-forming assays in these mice showed low abundance of the transduced human a-globin gene in their BFU-E and CFU-GEMM and the lack of its transcript. Thus, lentiviral vectors can be an effective vehicle for delivering the human α-globin gene into erythroid cells in utero, but, in the mouse model, delivery at late midgestation could not transduce hematopoietic stem cells adequately to sustain gene expression. © 2007 by The National Academy of Sciences of the USA. |
| Keywords: |
controlled study; human cell; genetics; nonhuman; protein localization; mouse; animal; metabolism; animals; mice; gene targeting; mus; reverse transcription polymerase chain reaction; gene expression; spleen; embryo; green fluorescent protein; cell line; animal experiment; animal model; vascularization; time; time factors; gene transfer; gene vector; genetic vectors; models, animal; gene expression regulation; prenatal development; gene expression regulation, developmental; liver; tissue distribution; messenger rna; rna, messenger; newborn; gene therapy; lentivirus; lentivirus vector; erythroid cell; transgene; reporter gene; pregnancy; genes, reporter; fetus; hematopoietic stem cell; animals, newborn; transgenes; yolk sac; lentivirinae; erythroid cells; gene transfer techniques; beta globin; southern blotting; alpha thalassemia; lentiviral vector; in utero gene transfer; yolk sac vessel injection; burst forming unit e; colony forming unit gemm; fetal therapy; alpha-thalassemia; fetal therapies
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