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Journal of Virology, February 2009, p. 2004-2010, Vol. 83, No. 4
0022-538X/09/$08.00+0     doi:10.1128/JVI.01298-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Herpes Simplex Virus Type 1 Thymidine Kinase Sequence Fused to the lacZ Gene Increases Levels of β-Galactosidase Activity per Genome of High-Capacity but Not First-Generation Adenoviral Vectors In Vitro and In Vivo{triangledown} ,{dagger}

M. Puntel,1 R. J. Barrett,1 S. Mondkar,1 V. Saxena,1 K. M. Kroeger,1 A. K. M. Muhammad,1 C. Liu,1 N. Bondale,1 S. Sciascia,1 W. Xiong,1 Y. Shi,1 A. Salem,1 A. Zadmehr,1 P. Huynh,1 D. Palmer,2 P. Ng,2 M. G. Castro,1 and P. R. Lowenstein1*

Board of Governors, Gene Therapeutics Research Institute, Cedars-Sinai Medical Center, and Departments of Medicine and Molecular and Medical Pharmacology, University of California Los Angeles, Los Angeles, California 90048,1 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 770302

Received 21 June 2008/ Accepted 20 November 2008

Increased transgene expression per vector genome is an important goal in the optimization of viral vectors for gene therapy. Herein we demonstrate that herpes simplex virus type 1 (HSV1) thymidine kinase (TK) gene sequences (1,131 bp) fused to the 3' end of lacZ increase transgene expression from high-capacity adenoviral vectors (HCAd), but not from first-generation (Ad) vectors. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), in contrast, increased transgene expression levels from Ad but not HCAd vectors. The differential activity of the HSV1 TK gene and WPRE sequences was detected both in vitro and in vivo and suggests potentially different mechanisms of action or the interaction of these elements with vector genomic sequences.

* Corresponding author. Mailing address: Board of Governors, Gene Therapeutics Research Institute, Davis Bldg., Room 5090, Cedars-Sinai Medical Center, 8700 Beverly Blvd., Los Angeles, CA 90048. Phone: (310) 423-7330. Fax: (310) 423-7308. E-mail: lowensteinp{at}cshs.org

{triangledown} Published ahead of print on 10 December 2008.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, February 2009, p. 2004-2010, Vol. 83, No. 4
0022-538X/09/$08.00+0     doi:10.1128/JVI.01298-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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