The DOI matches its Crossref record; 34 of the 34 references it cites were found in a registry or an index.
The woodchuck hepatitis virus post-transcriptional regulatory element reduces readthrough transcription from retroviral vectors
Abstract
The woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) increases transgene expression from a variety of viral vectors, although the precise mechanism is not known. WPRE is most effective when placed downstream of the transgene, proximal to the polyadenylation signal. We hypothesized that WPRE likely reduces viral mRNA readthrough transcription by improving transcript termination, which in turn would increase viral titers and expression. Using a Cre-lox-mediated plasmid-based assay, we found significant readthrough transcription from gamma-retroviral vector (RV) long terminal repeat (wt RV-LTR) and RV LTR with a self-inactivating deletion (SIN RV-LTR). WPRE, when placed upstream of the RV LTRs, significantly reduced readthrough transcription. Readthrough, present at much lower levels with the SIN HIV-1 LV-LTR, was also reduced with WPRE. When placed in RV vectors, WPRE increased total RV genomic mRNA; and increased viral titers from transiently transfected 293T cells and stable PG13 producer cells by 7- to 15-fold. The mechanism of increased titers and expression was not due to increased nuclear mRNA export, increased rate of viral transcription or a significant increase in viral mRNA half-life. Our results showed that WPRE improved vector genomic transcript termination to increase titers and expression from RVs.
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“The woodchuck hepatitis post-transcriptional regulatory element (WPRE) is often included in AAV cassettes to improve viral titer and transduction efficiency via stabilizing mRNA and regulating transcriptional termination [26, 27].”
From Designing and optimizing AAV-mediated gene therapy for neurodegenerative diseases: from bench to bedside (2024) · cites this paper as [27]
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Metadata
- Authors
- Higashimoto, T., Urbinati, F., Perumbeti, A., Jiang, G., Zarzuela, A., Chang, L.-J., Kohn, D. B., & Malik, P.
- Affiliations
- University of Southern California (US)
- University of Florida (US)
- Corresponding author
- Malik, P
- Venue
- Gene Therapy
- Published
- 2007-06-28
- Volume
- 14
- Issue
- 17
- Pages
- 1298-1304
- ISSN
- 0969-7128 (ISSN-L)
- 1476-5462
- OpenAlex
- W1975554944
- Cited by
- 101
- References
- 34
- Citations by year
- 2026: 13 · 2025: 9 · 2024: 10 · 2023: 2 · 2022: 10 · 2021: 4 · 2020: 3 · 2019: 3 · 2018: 1 · 2017: 6 · 2016: 3 · 2015: 2 · 2014: 6 · 2013: 3 · 2012: 5
- FWCI
- 2.86
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5u54hl070595-05)
- National Heart, Lung, and Blood Institute (P01HL073104, U54-HL070595, R01-HL70135-01, R01HL070135)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- read-through transcription, retroviral vectors, long terminal repeat, HIV-1 long terminal repeat, viral titer, transgene expression, polyadenylation signal
- MeSH terms
- Animals, Cell Line, Genetic Engineering, Genetic Vectors, Humans, Leukemia Virus, Murine, Retroviridae, RNA Processing, Post-Transcriptional, Transcription, Genetic, Transfection, Simian Immunodeficiency Virus, Gene Expression, Gene Expression Regulation, Viral, Reading Frames, Hepatitis B Virus, Woodchuck, Transgenes, Terminal Repeat Sequences, Regulatory Elements, Transcriptional
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