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Retroviral transfer of a chimeric multidrug resistance‐adenosine deaminase gene
Author(s) -
Germann Ursula A.,
Chin KhewVoon,
Pastan Ira,
Gottesman Michael M.
Publication year - 1990
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.4.5.1968408
Subject(s) - adenosine deaminase , multiple drug resistance , gene transfer , chemistry , adenosine , gene , virology , medicine , biology , drug resistance , microbiology and biotechnology , biochemistry
A fusion between a selectable multidrug resistance (MDR 1) cDNA and an adenosine deaminase (ADA) cDNA concomitantly confers multidrug resistance and ADA activity on transfected cells. We have produced a Harvey murine sarcoma virus‐derived, replication‐defective, recombinant retrovirus to transduce this chimeric MDR‐ADA gene efficiently into a great variety of cells. Infection with the MDR‐ADA retrovirus conferred the multidrug resistance phenotype on drug‐sensitive cells, therefore allowing selection in the presence of colchicine. Colchicine‐resistant cells synthesized large amounts of a membrane‐associated 210‐kDa MDR‐ADA fusion protein that preserved both MDR and ADA functional activities. To monitor expression of the chimeric gene in vivo, Kirsten virus‐transformed NIH cells were infected with the MDR‐ADA retrovirus, and after drug‐selection, injected into athymic nude mice. Tumors developed that contained the bifunctionally active MDR‐ADA fusion protein. When these mouse tumor cells were placed in tissue culture without the selecting drug, they did not lose the bifunctionally active MDR‐ADA fusion protein. The replication‐defective, recombinant MDR‐ADA retrovirus should be useful to stably introduce the chimeric MDR‐ADA gene into a variety of cell types for biological experiments in vitro and in vivo.— G ermann , U. A.; C hin , K.‐V.; P astan , I.; G ottesman , M. M. Retroviral transfer of a chimeric multidrug resistance‐adenosine deaminase gene. FASEB J. 4: 1501‐1507; 1990.