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Journal of Virology, July 2000, p. 6659-6668, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Novel Tat-Encoding Bicistronic Human Immunodeficiency Virus Type 1-Based Gene Transfer Vectors for High-Level Transgene Expression

Narasimhachar Srinivasakumar* and Friedrich Schuening

Division of Hematology-Oncology, Department of Medicine, Vanderbilt University, Nashville, Tennessee

Received 22 October 1999/Accepted 14 April 2000

We describe bicistronic single-exon Tat (72-amino-acid Tat [Tat72])- and full-length Tat (Tat86)-encoding gene transfer vectors based on human immunodeficiency virus type 1 (HIV-1). We created versions of these vectors that were rendered Rev independent by using the constitutive transport element (CTE) from Mason-Pfizer monkey virus (MPMV). Tat72-encoding vectors performed better than Tat86-expressing vectors in gene transfer experiments. CTE-containing vectors, produced in a Rev-independent packaging system, had gene transfer efficiencies nearly equivalent to those produced using a combination RNA transport (CTE and Rev-Rev response element)-based packaging system. The Tat72-encoding vectors could be efficiently transduced into a variety of cell types, showed higher levels of transgene expression than vectors with the simian cytomegalovirus immediate-early or the simian virus 40 early promoter, and provide an alternative to HIV-1 vectors with internal promoters.

* Corresponding author. Mailing address: 547 MRBII, 2220 Pierce Ave., Division of Hematology-Oncology, Department of Medicine, Vanderbilt University, Nashville, TN 37232-6305. Phone: (615) 936-2134. Fax: (615) 936-3853. E-mail: narasimhachar.srinivasakumar{at}mcmail.vanderbilt.edu.

Journal of Virology, July 2000, p. 6659-6668, Vol. 74, No. 14
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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