Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors Journal Article

  

Authors:	Li, M.; Hantzopoulos, P. A.; Banerjee, D.; Zhao, S. C.; Schweitzer, B. I.; Gilboa, E.; Bertino, J. R.
Article Title:	Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors
Abstract:	To determine the effect of different promoters on the expression of an altered dihydrofolate reductase (DHFR) gene conferring methotrexate (MTX) resistance in different cell types, double-copy retroviral vectors were constructed carrying a murine mutant DHFR under the control of five different promoters, i.e., human adenosine deaminase (ADA), simian virus 40 (SV40), thymidine kinase (TK), human β-actin, and cytomegalovirus (CMV). Their expression was compared in NIH-3T3 cells, three human leukemia cell lines, and mouse bone marrow. The variant DHFR is readily expressed from these various promoters in retroviral vectors at a selectable level. In 3T3 cells, the DHFR constructs containing the SV40 promoter conferred the highest levels of resistance to MTX. In K562 and Raji cells, the construct with the TK promoter produced the highest level of resistance. However granulocyte-macrophage colony-forming unit (CFU-GM) colonies from mouse marrow were more resistant to MTX when infected with vectors containing the SV40 promoter and ADA promoter as compared to the other promoter constructs. These studies show that, mouse fibroblast cell lines such as NIH-3T3 do not predict the effectiveness of retroviral-mediated gene transfer for marrow progenitor cells, and that the activity of retroviral vector-encoded promoters vary in an unpredictable manner from cell type to cell type. Possible implications for basic gene transfer studies and clinical applications are discussed.
Keywords:	leukemia; promoter region; genetics; mutation; comparative study; methotrexate; mouse; animal; metabolism; mice; bone marrow cells; drug resistance; tumor cells, cultured; genetic vectors; gene expression regulation; cell culture; murinae; simian virus 40; bone marrow cell; dihydrofolate reductase; tetrahydrofolate dehydrogenase; blotting, northern; moloney leukemia oncovirus; moloney murine leukemia virus; cytomegalovirus; cell strain 3t3; 3t3 cells; northern blotting; gene expression regulation, enzymologic; promoter regions (genetics); simiae; dna vector; simian virus; human; article; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; ada
Journal Title:	Human Gene Therapy
Volume:	3
Issue:	4
ISSN:	1043-0342
Publisher:	Mary Ann Liebert, Inc
Date Published:	1992-08-01
Start Page:	381
End Page:	390
Language:	English
DOI:	10.1089/hum.1992.3.4-381
PUBMED:	1525211
PROVIDER:	scopus
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0026903297&doi=10.1089%2fhum.1992.3.4-381&partnerID=40&md5=acd58c98869e059c0204dea07e0ce8c7
Notes:	Article -- Export Date: 30 July 2019 -- Source: Scopus

https://synapse.mskcc.org/synapse/works/has_related_data_chk/149741