Evaluation of the T-REx transcription switch for conditional expression and regulation of HSV-1 vectors

Virus Genes
Charles Werner KnopfJúlius Rajcáni

Abstract

Herpes simplex virus 1 (HSV-1) strain ANG and ANGpath were cloned as bacterial artificial chromosome (BAC). Two different types of BAC genomes were obtained. BAC genomes of type I contained the BAC replicon at the intended target region between the genes of UL48 and UL49. In BAC genomes of type II, the BAC sequences were found to be aberrantly fused between the termini of the HSV-1 genome. Both the BAC types were used to establish a conditional gene expression system for HSV-1 by Flp recombinase-mediated insertion of expression vectors that were modified to respond to the T-REx tetracycline (Tet)-inducible transcription switch. During BAC cloning and mutagenesis in E. coli, not only deletions but also defined mutations of the HSV-1 genome were observed. Successful virus reconstitution from BACs with large inserts demonstrated that HSV-1 has a packaging capacity for foreign sequences of at least 8.1% of its genome size. Targets for Tet-regulated gene expression were the viral DNA polymerase gene (pol) and a reporter gene of glycoprotein B fused to enhanced green fluorescent protein (gBGFP). Results with the pol gene as target showed that virus plaque production could not be significantly controlled by the T-REx gene switch using...Continue Reading

References

Jul 1, 1988·The Journal of General Virology·D J McGeochP Taylor
Feb 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·R R Spaete, N Frenkel
Nov 11, 1986·Nucleic Acids Research·C W KnopfH C Kaerner
Jun 14, 1967·Journal of Molecular Biology·B Hirt
Nov 15, 1996·The Journal of Biological Chemistry·F J Kühn, C W Knopf
Feb 7, 1998·Proceedings of the National Academy of Sciences of the United States of America·M MesserleU H Koszinowski
Jan 1, 1997·Intervirology·J R Berger, A Nath
Oct 15, 1998·Nature Genetics·Y ZhangA F Stewart
Apr 26, 2000·Proceedings of the National Academy of Sciences of the United States of America·G A Smith, L W Enquist
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Jul 11, 2001·Genome Génome / Conseil National De Recherches Canada·J SongJ Jiang
Feb 8, 2003·DNA and Cell Biology·Edward A BurtonJoseph C Glorioso
Mar 17, 2004·Methods in Molecular Biology·Eva-Maria BorstMartin Messerle
Dec 15, 2004·Journal of Virology·Brigitte RuppUlrich H Koszinowski
Feb 26, 2005·Journal of Virology·Eva-Maria Borst, Martin Messerle
Apr 1, 2006·Molecular Therapy : the Journal of the American Society of Gene Therapy·Feng YaoZheming Lu

❮ Previous
Next ❯

Citations

Jun 8, 2021·Virology : Research and Treatment·Prapti H ModyJuliet V Spencer

❮ Previous
Next ❯

Related Concepts

Related Feeds

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.