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

Ai-Li Shiau, H. M. Yang, P. Wu, Chao-Liang Wu

Research output: Contribution to journalArticle

9 Citations (Scopus)

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 provide 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.

Original languageEnglish
Pages (from-to)1571-1574
Number of pages4
JournalGene Therapy
Volume5
Issue number11
DOIs
Publication statusPublished - 1998 Jan 1

Fingerprint

Cytosine Deaminase
Flucytosine
Genes
Fluorouracil
Uracil
Cytosine
Transgenes
Poisons
Catalysis
Genetic Therapy
Suicide

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

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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 provide 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.",
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Provision of positive and negative selections in retroviral vectors containing the cytosine deaminase gene. / Shiau, Ai-Li; Yang, H. M.; Wu, P.; Wu, Chao-Liang.

In: Gene Therapy, Vol. 5, No. 11, 01.01.1998, p. 1571-1574.

Research output: Contribution to journalArticle

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