This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Vanin, E. F. Articles by Jane, S. M. Search for Related Content PubMed PubMed Citation Articles by Vanin, E. F. Articles by Jane, S. M.

 Previous Article  |  Next Article 

J. Virol., 10 1997, 7820-7826, Vol 71, No. 10
Copyright © 1997, American Society for Microbiology

Development of high-titer retroviral producer cell lines by using Cre- mediated recombination

EF Vanin, L Cerruti, N Tran, G Grosveld, JM Cunningham and SM Jane
Rotary Bone Marrow Research Laboratory, Royal Melbourne Hospital Research Foundation, Parkville, Victoria, Australia.

Retroviral gene transfer is widely used in experimental and human gene therapy applications. We have devised a novel method of generating high- titer retroviral producer cell lines based on the P1 bacteriophage recombinase system Cre-loxP. Incorporation of loxP sites flanking a Neo(r)-SVTK cassette in the proviral DNA allows excision of these selectable markers through expression of Cre recombinase after production of a high-titer producer cell line. The resultant producer line contains a single loxP site flanked by the viral long terminal repeats. Retransfection of this line with the Cre expression vector and a plasmid containing a gene of interest flanked by loxP sites allows insertional recombination of the gene into the favorable preexisting site in the genome and the generation of a new line with a titer equivalent to that of the parental producer cell line. The efficiency of the process is sufficient to allow the generation of multiple new producer lines without the addition of antibiotic resistance genes. We have successfully generated retroviral vectors carrying different genes by using this approach and discuss the potential applications of this method in gene therapy.

This article has been cited by other articles:

• Satoh, W., Hirai, Y., Tamayose, K., Shimada, T. (2000). Site-Specific Integration of an Adeno-Associated Virus Vector Plasmid Mediated by Regulated Expression of Rep Based on Cre-loxP Recombination. J. Virol. 74: 10631-10638 [Abstract] [Full Text]  
• Darcy, P. K., Haynes, N. M., Snook, M. B., Trapani, J. A., Cerruti, L., Jane, S. M., Smyth, M. J. (2000). Redirected Perforin-Dependent Lysis of Colon Carcinoma by Ex Vivo Genetically Engineered CTL. J. Immunol. 164: 3705-3712 [Abstract] [Full Text]