High-level erythroid lineage-directed gene expression using globin gene regulatory elements after lentiviral vector-mediated gene transfer into primitive human and murine hematopoietic cells.
Lentiviral vectors efficiently transduce primitive human hematopoietic cells and are capable of transferring complex genomes. Vectors were designed with hypersensitive sites containing regulatory elements from the beta-globin locus control region linked to the beta-globin gene promoter to drive expression of the enhanced green fluorescent protein marker to facilitate analysis of the pattern of gene expression in various hematopoietic lineages. Such vectors gave higher level, induced expression in mouse erythroleukemia cells than a previously described vector that utilized an enhancer from the alpha locus and the ankyrin-1 promoter [Moreau-Gaudry, F., Xia, P., Jiang, G., Perelman, N.P., Bauer, G., Ellis, J., Surinya, K.H., Mavilio, F., Shen, C.K., and Malik, P. (2001). Blood 98, 2664-2672]. The addition of gamma-globin intron sequences further augmented vector expression. Expression was also effectively targeted to the erythroid lineage in cultured human cells from peripheral blood and in mouse red blood cells in vivo, although lower levels of expression were also observed in other lineages. Thus, these newly described vectors provide a means to achieve high-level gene expression, predominantly in erythroid cells, an outcome that may have potential therapeutic application.
Hanawa
Hideki
H
Division of Experimental Hematology, St. Jude Children's Research Hospital, 332 N Lauderdale, Memphis, TN 38105, USA.
Persons
Derek A
DA
Nienhuis
Arthur W
AW
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics; D001854:Bone Marrow Cells / Q000378:metabolism; D016026:Bone Marrow Transplantation; D019070:Cell Lineage; D004742:Enhancer Elements, Genetic; D015672:Erythroid Precursor Cells / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005809:Genes, Regulator; D005822:Genetic Vectors / Q000235:genetics; D005914:Globins / Q000235:genetics*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000821:virology; D006801:Humans; D007438:Introns; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D018952:Antigens, CD34; D008164:Luminescent Proteins; D016179:Granulocyte Colony-Stimulating Factor; D049452:Green Fluorescent Proteins; D005914:Globins