Abstract
We have studied the regulation of the human ß-globin gene after retroviral transfer into a variety of transformed and normal hematopoietic cells. After transfer into murine erythroleukemia cells (MEL) expression from the human ß-globin gene responds to inducers of erythroid maturation in parallel to the endogenous murine globin genes. After infection of human BFU-E, RNA expression from the virally-transferred ß-globin gene was measured at 2.57o-57o of the endogenous ß-globin level. The most improved globin vectors can transfer the human ß-globin gene into pluripotent hematopoietic stem cells in mouse bone marrow. Mice reconstituted with infected marrow show human ß-globin RNA and protein expression in peripheral blood cells for over 4 months. In these animals, both myeloid and lymphoid cells carry the integrated provirus at a level of about 1 copy per cell. In serial transplantation experiments, bone marrow from these animals is capable of repopulating secondary and tertiary recipient animals which go on to show long-term human ß-globin expression. Retroviral vectors thus provide a practical way to refine models of globin gene regulation through in vivo tests and to evaluate the feasibility of protocols for gene addition therapy.
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References
Behringer R.R., R.E. Hammer, R.L. Brinster, R.D. Palmiter, T.M. Townes. 1987. Two 3’ sequences direct erythroid-specific expression of human 3-globin genes in trangenic mice. Proc. Natl. Acad. Sci. USA 84: 7056–7060.
Bender, M.A., A.D. Miller, R.E. Gelinas. 1988. Expression of the human 3-globin gene after retroviral transfer into murine erythroleukemia cells and human BFU-E cells. Mol. Cell Biol. 8: 1725–1735
Bender, M.A., R.E. Gelinas, A.D. Miller. 1989. A majority of mice show long-term expression of a human 3-globin gene after retroviral transfer into hematopoietic stem cells. Mol. Cell Biol. 9: 1426–1434.
Bradshaw, H.D., W.W. Parson, M. Sheffer, P.J. Lioubin, E.R. Mulvihill, M.P. Gordon. 1987. Design, construction, and use of an electro-porator for plant protoplasts and animal cells. Anal. Biochem. 166: 342–348.
Brinster, R.L., J.M. Allen, R.R. Behringer, R.E. Gelinas, R.D. Palmiter. 1988. Introns increase transcriptional efficiency in transgenic mice. Proc. Natl Acad. Sci. USA 85: 836–840.
Chu, G., H. Hayakawa, P. Berg. 1987. Electroporation for the efficient transfection of mammalian cells with DNA. Nuc. Acids Res. 15: 1311–1326.
Cone, R.D., A.W. Benarous, D. Baorto, R.C. Mulligan. 1987. Regulated expression of a complete human 3-globin gene encoded by a transmissible retrovirus vector. Mol. Cell Biol. 7: 887–897.
Dick, J.E., M.C. Magli, D. Huszar, R.A. Phillips, A. Bernstein. 1985. Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hematopoietic system of W/Wv mice. Cell 42: 71–79.
Dzierzak, E.A., T. Papayannopoulou, R.C. Mulligan. 1988. Lineage-specific expression of a human 3-globin gene in murine bone marrow transplant recipient reconstituted with retrovirus-transduced stem cells. Nature 331: 35–41.
Forrester, W.C., S. Takegawa, T. Papayannopoulou, G. Stamatoyannopoulos, M. Groudine. 1987. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nuc. Acids Res. 15: 10159 – 10177.
Forrester, W.C., S. Takegawa, T. Papayannopoulou, G. Stamatoyannopoulos, M. Groudine. 1987. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nuc. Acids Res. 15: 10159–10177.
Forrester, W.C., S. Takegawa, T. Papayannopoulou, G. Stamatoyannopoulos, M. Groudine. 1987. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nuc. Acids Res. 15: 10159–10177.
Grosveld, F., G.B. van Assendelft, D.R. Greaves, G. Kollias. 1987. Position-independent, high level expression of the human ß-globin gene in transgenic mice. Cell 51: 975–985.
Harrison, D.E., CM. Astle. 1976. Population of lymphoid tissues in cured W-anemic mice by donor cells. Transplantation 22: 42–46.
Harrison, D.E., CM. Astle, J.A. DeLaittre. 1979. Processing by the thymus is not required for cells that cure and populate W/Wv recipients. Blood 54: 1152–1157.
Karlsson, S., T. Papayannopoulou, S.G. Schweiger, G. Stamatoyannopoulos, A.W. Nienhuis. 1987. Retroviral-mediated transfer of genomic globin genes leads to regulated production of RNA and protein. Proc. Natl. Acad. Sci. USA 84: 2411–2415.
Karlsson, S., D.M. Bodine, L. Perry, T. Papayannopoulou, A.W. Nienhuis. 1988. Expression of the human 3-globin gene following retroviral-mediated transfer into multipotential hematopoietic progenitorsof mice. Proc. Natl. Acad. Sci. USA 85: 6062–6066.
Keller, G., P. Paige, E. Gilboa, E.F. Wagner. 1985. Expressionof a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature 318: 149– 154.
Lemischka, I.R., D.H. Raulet, R.C. Mulligan. 1986. Developmental potential and dynamic behavior of hematopoietic stem cells. Cell 45: 917–927.
Lerner, N., S. Brigham, S. Goff, A. Bank. 1987. Human 3-globin expression after gene transfer using retroviral vectors. DNA 6: 573–582.
Miller, A.D. 1989. Retroviral Vectors. In: Current Topics in Microbiology and Immunology. (N. Muzyczka, Ed.) (In Press).
Miller, A.D., E.S. Ong, M.G. Rosenfeld, I.M. Verma, R.M. Evans. 1984. Infectious and selectable retrovirus containing an inducible rat growth hormone minigene. Science 225: 993 – 998.
Miller, A.D., C. Buttimore. 1986. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol. Cell Biol. 6: 2895–2902.
Miller, A.D., M.A. Bender, E.A.S. Harris, M. Kaleko, R.E. Gelinas. 1988. Design of retroviral vectors for transfer and expression of the human 3 globin gene. J. Virol. 62: 4337–4345.
Trudel, M., F. Costantini. 1987. A 3’ enhancer contributes to the stage-specific expression of the human (3-globin gene. Genes Dev. 1: 954–961.
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© 1989 Plenum Press, New York
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Gelinas, R.E., Bender, M.A., Miller, A.D., Novak, U. (1989). Long-Term Expression of the Human ß-Globin Gene After Retroviral Transfer into Pluripotent Hematopoietic Stem Cells of the Mouse. In: Ascensao, J.L., Zanjani, E.D., Tavassoli, M., Levine, A.S., MacKintosh, F.R. (eds) Molecular Biology of Erythropoiesis. Advances in Experimental Medicine and Biology, vol 271. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-0623-8_15
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DOI: https://doi.org/10.1007/978-1-4613-0623-8_15
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