Novel Pol II Fusion Promoter Directs Human Immunodeficiency Virus Type 1-inducible Coexpression of a Short Hairpin RNA and Protein

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2006 Jan 28

PMID 16439542

Citations 27

Authors

Hoshang J Unwalla

Hai-Tang Li

Ingrid Bahner

Ming-Jie Li

Donald Kohn

John J Rossi

Affiliations

Soon will be listed here.

Abstract

We demonstrate a novel approach for coexpression of a short hairpin RNA (shRNA) with an open reading frame which exploits transcriptional read-through of a minimal polyadenylation signal from a Pol II promoter. We first observed efficient inducible expression of enhanced green fluorescent protein along with an anti-rev shRNA. We took advantage of this observation to test coexpression of the transdominant negative mutant (humanized) of human immunodeficiency type 1 (HIV-1) Rev (huRevM10) along with an anti-rev shRNA via an HIV-1-inducible fusion promoter. The coexpression of the shRNA and transdominant protein resulted in potent, long-term inhibition of HIV-1 gene expression and suppression of shRNA-resistant mutants. This dual expression system has broad-based potential for other shRNA applications, such as cases where simultaneous knockdown of mutant and wild-type transcripts must be accompanied by replacement of the wild-type protein.

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