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High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector
Tissue-/cancer-specific promoters for use in adenovirus vectors (AdVs) are valuable for elucidating specific gene functions and for use in gene therapy. However, low activity, non-specific expression and size limitations in the vector are always problems. Here, we developed a ‘double-unit’ AdV conta...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3025582/ https://www.ncbi.nlm.nih.gov/pubmed/21051352 http://dx.doi.org/10.1093/nar/gkq966 |
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author | Kanegae, Yumi Terashima, Miho Kondo, Saki Fukuda, Hiromitsu Maekawa, Aya Pei, Zheng Saito, Izumu |
author_facet | Kanegae, Yumi Terashima, Miho Kondo, Saki Fukuda, Hiromitsu Maekawa, Aya Pei, Zheng Saito, Izumu |
author_sort | Kanegae, Yumi |
collection | PubMed |
description | Tissue-/cancer-specific promoters for use in adenovirus vectors (AdVs) are valuable for elucidating specific gene functions and for use in gene therapy. However, low activity, non-specific expression and size limitations in the vector are always problems. Here, we developed a ‘double-unit’ AdV containing the Cre gene under the control of an α-fetoprotein promoter near the right end of its genome and bearing a compact ‘excisional-expression’ unit consisting of a target cDNA ‘upstream’ of a potent promoter between two loxPs near the left end of its genome. When Cre was expressed, the expression unit was excised as a circular molecule and strongly expressed. Undesired leak expression of Cre during virus preparation was completely suppressed by a dominant-negative Cre and a short-hairpin RNA against Cre. Using this novel construct, a very strict specificity was maintained while achieving a 40- to 90-fold higher expression level, compared with that attainable using a direct specific promoter. Therefore, the ‘double-unit’ AdV enabled us to produce a tissue-/cancer-specific promoter in an AdV with a high expression level and strict specificity. |
format | Text |
id | pubmed-3025582 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-30255822011-01-24 High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector Kanegae, Yumi Terashima, Miho Kondo, Saki Fukuda, Hiromitsu Maekawa, Aya Pei, Zheng Saito, Izumu Nucleic Acids Res Methods Online Tissue-/cancer-specific promoters for use in adenovirus vectors (AdVs) are valuable for elucidating specific gene functions and for use in gene therapy. However, low activity, non-specific expression and size limitations in the vector are always problems. Here, we developed a ‘double-unit’ AdV containing the Cre gene under the control of an α-fetoprotein promoter near the right end of its genome and bearing a compact ‘excisional-expression’ unit consisting of a target cDNA ‘upstream’ of a potent promoter between two loxPs near the left end of its genome. When Cre was expressed, the expression unit was excised as a circular molecule and strongly expressed. Undesired leak expression of Cre during virus preparation was completely suppressed by a dominant-negative Cre and a short-hairpin RNA against Cre. Using this novel construct, a very strict specificity was maintained while achieving a 40- to 90-fold higher expression level, compared with that attainable using a direct specific promoter. Therefore, the ‘double-unit’ AdV enabled us to produce a tissue-/cancer-specific promoter in an AdV with a high expression level and strict specificity. Oxford University Press 2011-01 2010-11-04 /pmc/articles/PMC3025582/ /pubmed/21051352 http://dx.doi.org/10.1093/nar/gkq966 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methods Online Kanegae, Yumi Terashima, Miho Kondo, Saki Fukuda, Hiromitsu Maekawa, Aya Pei, Zheng Saito, Izumu High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title | High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title_full | High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title_fullStr | High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title_full_unstemmed | High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title_short | High-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
title_sort | high-level expression by tissue/cancer-specific promoter with strict specificity using a single-adenoviral vector |
topic | Methods Online |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3025582/ https://www.ncbi.nlm.nih.gov/pubmed/21051352 http://dx.doi.org/10.1093/nar/gkq966 |
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