In Situ Hybridization for Detection of AAV-Mediated Gene Expression.

In Situ Hybridization for Detection of AAV-Mediated Gene Expression. Methods Mol Biol. 2019;1950:107-122 Authors: Hunter JE, Gurda BL, Yoon SY, Castle MJ, Wolfe JH Abstract Techniques to localize vector transgenes in cells and tissues are essential in order to fully characterize gene therapy outcomes. In situ hybridization (ISH) uses synthesized complementary RNA or DNA nucleotide probes to localize and detect sequences of interest in fixed cells, tissue sections, or whole tissue mounts. Variations in techniques include adding labels to probes, such as fluorophores, which can allow for the simultaneous visualization of multiple targets. Here we provide the steps necessary to: (1) label probes for colorimetric visualization and (2) perform ISH on OCT cryo-preserved fixed frozen tissues. PMID: 30783970 [PubMed - indexed for MEDLINE]
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research