Provision of positive and negative selections in retroviral vectors containing the cytosine deaminase gene

Shiau, A-L; Yang, H-M; Wu, P; Wu, C-L

Abstract

The E. coli cytosine deaminase (CD) provides a negative selection system for suicide gene therapy as CD transfectants are eliminated following 5-fluorocytosine (5FC) treatment. Here we report a positive selection system for the CD gene using 5-fluorouracil (5FU) and cytosine in selection medium to screen for CD-positive transfectants. It is based on the relief of 5FU toxicity by uracil which is converted from cytosine via CD catalysis, as uracil competes with the toxic 5FU in subsequent pyrimidine metabolism. Hence, a retroviral vector containing the CD gene may pro- vide both positive and negative selections after gene transfer. The CD transfectants selected with the positive selection system showed susceptibility to 5FC in subsequent negative selection in vitro and in vivo. Therefore, this dual selection system is useful not only for combination therapy with transgene and CD gene, but can also act to eliminate selectively transduced cells after the transgene has furnished its effects or upon undesired conditions if 5FC is applied for negative selection in vivo.

Citation

Shiau, A., Yang, H., Wu, P., & Wu, C. (1998). Provision of positive and negative selections in retroviral vectors containing the cytosine deaminase gene. Gene Therapy, 5, 1571–1574. https://doi.org/10.1038/sj.gt.3300757

• Journal Article Type: Article
• Online Publication Date: Nov 13, 1998
• Publication Date: 1998-11
• Deposit Date: Apr 19, 2024
• Journal: Gene Therapy
• Print ISSN: 0969-7128
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 5
• Pages: 1571–1574
• DOI: https://doi.org/10.1038/sj.gt.3300757
• Publisher URL: https://www.nature.com/articles/3300757