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

Autor: Hunter JE; Research Institute of the Children's Hospital of Philadelphia, Philadelphia, PA, USA., Gurda BL; Research Institute of the Children's Hospital of Philadelphia, Philadelphia, PA, USA., Yoon SY; Research Institute of the Children's Hospital of Philadelphia, Philadelphia, PA, USA.; W.F. Goodman Center for Comparative Medical Genetics, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA., Castle MJ; Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA., Wolfe JH; Research Institute of the Children's Hospital of Philadelphia, Philadelphia, PA, USA. jhwolfe@vet.upenn.edu.; W.F. Goodman Center for Comparative Medical Genetics, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA. jhwolfe@vet.upenn.edu.; Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jhwolfe@vet.upenn.edu.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2019; Vol. 1950, pp. 107-122.
DOI: 10.1007/978-1-4939-9139-6_6
Abstrakt: 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.
Databáze: MEDLINE