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Transfer of Murine Leukaemia and Murine Sarcoma Virus Genetic Information by Transfection with Isolated Metaphase Chromosomes

¹ Laboratory of Molecular Genetics
and² Pregnancy Research Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.
³ Biological Carcinogenesis Program, Frederick Cancer Research Facility, Frederick, Maryland 21701, U.S.A.
and⁴ Dermatology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205, U.S.A.

Chromosome-mediated transfer of murine leukaemia (MuLV) and murine sarcoma (MuSV) virus genetic information to uninfected recipient cells was investigated. Metaphase chromosomes from AKR MuLV-infected SC-1 mouse cells were incubated with NIH/3T3 cells. After several passages (1 to 3 weeks), infectious virions exhibiting reverse transcriptase activity and the characteristic host range of ecotropic, N-tropic AKR virus appeared in the supernatant fluids of the treated cells. Restriction endonuclease analysis of genomic DNA from transfected cells indicated that AKR proviral DNA was associated with the high molecular weight DNA of the host. These results demonstrate that the AKR MuLV genome can be stably transferred to uninfected recipient cells via isolated metaphase chromosomes. Although AKR virions are not able to infect heterologous cells, chromosome-mediated transfection resulted in the establishment of productive AKR MuLV infection in mink cells. Thus, the use of chromosomes to transfer virus genes can circumvent the natural host restriction barrier. In other experiments, it was shown that normal NIH/3T3 cells were transformed after exposure to metaphase chromosomes isolated from an MuSV-infected, non-producer line. Foci were detected 14 to 21 days after chromosome treatment and were shown to contain true viral transformants since transforming virus was produced after superinfection with MuLV.

Keywords: murine retroviruses, metaphase chromosomes, gene transfer, transformation

Received 27 April 1982; accepted 4 June 1982.