ArticlePloS one2016
Three-Dimensional Imaging of the Intracellular Fate of Plasmid DNA and Transgene Expression: ZsGreen1 and Tissue Clearing Method CUBIC Are an Optimal Combination for Multicolor Deep Imaging in Murine Tissues.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- EBDE clearing: A strategy for simultaneous 3D visualization and quantitative analysis of both renal arteries and glomeruli.Theranostics · 2026Article
- A non-viral DNA delivery system consisting of multifunctional chimeric peptide fused with zinc-finger protein.iScience · 2024Article
- An optimized visualization and quantitative protocol for in-depth evaluation of lymphatic vessel architecture in the liver.American journal of physiology. Gastrointestinal and liver physiology · 2023Article
- Hydrodynamic Delivery: Characteristics, Applications, and Technological Advances.Pharmaceutics · 2023Review
- Understanding In Vivo Fate of Nucleic Acid and Gene Medicines for the Rational Design of Drugs.Pharmaceutics · 2021Review
- Suppression of Peritoneal Fibrosis by Sonoporation of Hepatocyte Growth Factor Gene-Encoding Plasmid DNA in Mice.Pharmaceutics · 2021Article
- A pH-Adjustable Tissue Clearing Solution That Preserves Lipid Ultrastructures: Suitable Tissue Clearing Method for DDS Evaluation.Pharmaceutics · 2020Article
- Biomedical Applications of Tissue Clearing and Three-Dimensional Imaging in Health and Disease.iScience · 2020Review
- Nanobubble Mediated Gene Delivery in Conjunction With a Hand-Held Ultrasound Scanner.Frontiers in pharmacology · 2020Article
- Monitoring method for transgene expression in target tissue by blood sampling.Biotechnology reports (Amsterdam, Netherlands) · 2019Article
- Looking through Brains with Fast Passive CLARITY: Zebrafish, Rodents, Non-human Primates and Humans.Bio-protocol · 2019Article
- Application of Direct Sonoporation from a Defined Surface Area of the Peritoneum: Evaluation of Transfection Characteristics in Mice.Pharmaceutics · 2019Article
- Whole-body clearing, staining and screening of calcium deposits in the mdx mouse model of Duchenne muscular dystrophy.Skeletal muscle · 2018Article
- CLARITY for High-resolution Imaging and Quantification of Vasculature in the Whole Mouse Brain.Aging and disease · 2018Article
- Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney.Journal of visualized experiments : JoVE · 2018Article
- Efficient gene transfection to the brain with ultrasound irradiation in mice using stabilized bubble lipopolyplexes prepared by the surface charge regulation method.International journal of nanomedicine · 2018Article
- Effective intraperitoneal gene transfection system using nanobubbles and ultrasound irradiation.Drug delivery · 2017Article
- Characterization of transgene expression and pDNA distribution of the suctioned kidney in mice.Drug delivery · 2017Article
- CUBIC pathology: three-dimensional imaging for pathological diagnosis.Scientific reports · 2017Article
- Kidney-specific transposon-mediated gene transfer in vivo.Scientific reports · 2017Article
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Evaluation methods for determining the distribution of transgene expression in the body and the in vivo fate of viral and non-viral vectors are necessary for successful development of in vivo gene delivery systems. Here, we evaluated the spatial distribution of transgene expression using tissue clearing methods. After hydrodynamic injection of plasmid DNA into mice, whole tissues were subjected to tissue clearing. Tissue clearing followed by confocal laser scanning microscopy enabled evaluation of the three-dimensional distribution of transgene expression without preparation of tissue sections. Among the tested clearing methods (ClearT2, SeeDB, and CUBIC), CUBIC was the most suitable method for determining the spatial distribution of transgene expression in not only the liver but also other tissues such as the kidney and lung. In terms of the type of fluorescent protein, the observable depth for green fluorescent protein ZsGreen1 was slightly greater than that for red fluorescent protein tdTomato. We observed a depth of ~1.5 mm for the liver and 500 μm for other tissues without preparation of tissue sections. Furthermore, we succeeded in multicolor deep imaging of the intracellular fate of plasmid DNA in the murine liver. Thus, tissue clearing would be a powerful approach for determining the spatial distribution of plasmid DNA and transgene expression in various murine tissues.
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