Novel Pol II fusion promoter directs human immunodeficiency virus type 1-inducible coexpression of a short hairpin RNA and protein

Hoshang J. Unwalla, Hai Tang Li, Ingrid Bahner, Ming Jie Li, Donald Kohn, John J. Rossi

Research output: Contribution to journalArticle

42 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1863-1873
Number of pages11
JournalJournal of Virology
Volume80
Issue number4
DOIs
StatePublished - Feb 1 2006

Fingerprint

small interfering RNA
Human immunodeficiency virus 1
Small Interfering RNA
HIV-1
promoter regions
Proteins
proteins
mutants
Polyadenylation
immunosuppression
green fluorescent protein
Open Reading Frames
open reading frames
Gene Expression
gene expression
testing

ASJC Scopus subject areas

  • Immunology

Cite this

Novel Pol II fusion promoter directs human immunodeficiency virus type 1-inducible coexpression of a short hairpin RNA and protein. / Unwalla, Hoshang J.; Li, Hai Tang; Bahner, Ingrid; Li, Ming Jie; Kohn, Donald; Rossi, John J.

In: Journal of Virology, Vol. 80, No. 4, 01.02.2006, p. 1863-1873.

Research output: Contribution to journalArticle

Unwalla, Hoshang J. ; Li, Hai Tang ; Bahner, Ingrid ; Li, Ming Jie ; Kohn, Donald ; Rossi, John J. / Novel Pol II fusion promoter directs human immunodeficiency virus type 1-inducible coexpression of a short hairpin RNA and protein. In: Journal of Virology. 2006 ; Vol. 80, No. 4. pp. 1863-1873.
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