Detection of Reporter Gene Expression in Murine Airways
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We have shown that to overcome the low levels of expression from gene transfer vector-mediated β-galactosidase expression in lung, it is essential to replace the cytoplasmic β-galactosidase gene with a nuclear targeted β-galactosidase gene. We found that lung should be sectioned and fixed prior to staining for β-galactosidase expression and that en bloc staining of intact lung is inefficient at staining positively transduced cells located deeper in the lung spaces. For GFP fluorescence, it is important to inflate the lungs with fixative prior to freezing and subsequent sectioning. For processing of nasal tissues for β-galactosidase expression, we expand on a protocol used in previously reported gene transfer studies.
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- Detection of Reporter Gene Expression in Murine Airways
- Book Title
- Reporter Genes
- Book Subtitle
- A Practical Guide
- pp 25-34
- Print ISBN
- Online ISBN
- Series Title
- Methods in Molecular Biology
- Series Volume
- Series ISSN
- Humana Press
- Copyright Holder
- Humana Press Inc.
- Additional Links
- LacZ, β-galactosidase
- placental alkaline phosphatase
- gene expression
- Industry Sectors
- Donald S. Anson (1)
- Editor Affiliations
- 1. Department of Genetic Medicine, Women’s and Children’s Hospital
- Author Affiliations
- 2. Department of Pathology, Gene Therapy Program, USA
- 3. Laboratory Medicine, University of Pennsylvania, USA
- 4. Gene Therapy Program, Department of Pathology and Laboratory Medicine, University of Pennsylvania, USA
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