PROVISION OF POSITIVE AND NEGATIVE SELECTIONS IN RETROVIRAL VECTORS CONTAINING THE CYTOSINE DEAMINASE GENE

Citation
Al. Shiau et al., PROVISION OF POSITIVE AND NEGATIVE SELECTIONS IN RETROVIRAL VECTORS CONTAINING THE CYTOSINE DEAMINASE GENE, Gene therapy, 5(11), 1998, pp. 1571-1574
Citations number
21
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biothechnology & Applied Migrobiology","Genetics & Heredity",Biology,"Medicine, Research & Experimental","Pharmacology & Pharmacy
Journal title
ISSN journal
0969-7128
Volume
5
Issue
11
Year of publication
1998
Pages
1571 - 1574
Database
ISI
SICI code
0969-7128(1998)5:11<1571:POPANS>2.0.ZU;2-I
Abstract
The E. coli cytosine deaminase (CD) provides a negative selection syst em for suicide gene therapy as CD transfectants are eliminated followi ng 5-fluorocytosine (5FC) treatment. Here we report a positive selecti on system for the CD gene using 5-fluorouracil (5FU) and cytosine in s election medium to screen for CD-positive transfectants. It is based o n the relief of 5FU toxicity by uracil which is converted form cytosin e via CD catalysis, as uracil competes with the toxic 5FU in subsequen t pyrimidine metabolism. Hence, a retroviral vector containing the CD gene may provide both positive and negative selections after gene tran sfer. The CD transfectants selected with the positive selection system showed susceptibility to 5FC in subsequent negative selection in vitr o and in vivo, Therefore, this dual I selection system is useful not o nly for combination therapy with transgene and CD gene, but can also a ct to eliminate, Selectively transduced cells after the transgene has furnished its effects or upon undesired conditions if 5FC is applied f or negative selection in vivo.