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Science 20 December 1985:
Vol. 230. no. 4732, pp. 1395 - 1398
DOI: 10.1126/science.2999985
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Articles
Science, Vol 230, Issue 4732, 1395-1398
Copyright © 1985 by American Association for the Advancement of Science
Gene expression in mice after high efficiency retroviral-mediated gene transfer
MA Eglitis,
P Kantoff,
E Gilboa,
and
WF Anderson
A retroviral expression vector (N2) containing the selectable gene, neoR, has been used to determine the optimal conditions for infecting murine hematopoietic progenitor cells at high efficiency. After infected bone marrow cells were introduced into lethally irradiated mice, the presence, stability, and expression of the vector DNA sequences were analyzed either in individual spleen foci 10 days later or in the blood, bone marrow, and spleens of mice 4 months later. When bone marrow cells were cultured in medium containing virus with titers of more than 10(6) colony-forming units per milliliter in the presence of purified murine interleukin-3, more than 85 percent of the resulting foci contained vector DNA. This proviral vector DNA was intact. Efficient expression of the neoR gene was demonstrated in most of the DNA-positive foci examined. The spleens of reconstituted animals (over a long term) contained intact "vector DNA" and the blood and bone marrow expressed the neoR gene in some animals. Thus, a retroviral vector can be used to introduce intact exogenous DNA sequences into hematopoietic stem cells with high efficiency and with substantial expression.
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