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Czech Academy of Sciences

4 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice
Takahiro Tadokoro 1, Atsushi Miyanohara 1, Michael Navarro 1, Kota Kamizato 1, Stefan Juhas 2, Jana Juhasova 2, Silvia Marsala 1, Oleksandr Platoshyn 1, Erik Curtis 3, Brandon Gabel 3, Joseph Ciacci 3, Nada Lukacova 4, Katarina Bimbova 4, Martin Marsala 1,4
1Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego, 2Institute of Animal Physiology and Genetics, Czech Academy of Sciences, 3Department of Neurosurgery, University of California, San Diego, 4Institute of Neurobiology, Slovak Academy of Sciences

The goal of the present study was to develop and validate the potency and safety of spinal adeno-associated virus 9 (AAV9)-mediated gene delivery by using a novel subpial gene delivery technique in adult mice.

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Biology

Practical Use of RNA Interference: Oral Delivery of Double-stranded RNA in Liposome Carriers for Cockroaches
Jia-Hsin Huang 1, Yun Liu 1, Yu-Hsien Lin 1,2,3, Xavier Belles 4, How-Jing Lee 1
1Department of Entomology, National Taiwan University, 2Biology Centre, Institute of Entomology, Czech Academy of Sciences, 3Faculty of Science, University of South Bohemia, 4Institute of Evolutionary Biology, CSIC-UPF

This manuscript demonstrates the depletion of gene expression in the midgut of the German cockroach through oral ingestion of double-stranded RNA encapsulated in liposomes.

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Biochemistry

Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis
Tomáš Zavřel 1, Dominik Chmelík 1,2, Maria A. Sinetova 3, Jan Červený 1
1Department of Adaptive Biotechnologies, Global Change Research Institute, Czech Academy of Sciences, 2Department of Plant Physiology, Faculty of Science, Masaryk University, 3Laboratory of Intracellular Regulation, Institute of Plant Physiology, Russian Academy of Sciences

Here, we present a protocol to quantitatively determine phycobiliprotein content in the cyanobacterium Synechocystis using a spectrophotometric method. The extraction procedure was also successfully applied to other cyanobacteria and algae strains; however, due to variations in pigment absorption spectra, it is necessary to test the spectrophotometric equations for each strain individually.

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Bioengineering

Subretinal Implantation of RPE on a Carrier in Minipigs: Guidelines for Preoperative Preparations, Surgical Techniques, and Postoperative Care
Lyubomyr Lytvynchuk *1,2, Zbynek Stranak *3, Hana Studenovska 4, David Rais 4, Štěpán Popelka 4, Lucie Tichotová 5,6, Yaroslav Nemesh 5,6, Anastasiia Kolesnikova 5,6, Ruslan Nyshchuk 5,6, Anna Brymová 5,6, Zdeňka Ellederová 5, Jana Čížková 5, Jana Juhásová 5, Štefan Juhás 5, Pavla Jendelová 7, Richárd Nagymihály 8, Igor Kozak 9, Slaven Erceg 10, Susanne Binder 11, Brigitte Müller 1, Knut Stieger 1, Jan Motlik 5, Goran Petrovski *8, Taras Ardan *5
1Eye Clinic, Department of Ophthalmology, University Hospital Giessen and Marburg GmbH, 2Karl Landsteiner Institute for Retinal Research and Imaging, 3Department of Ophthalmology, University Hospital Kralovske Vinohrady and Third Faculty of Medicine, Charles University in Prague, 4Institute of Macromolecular Chemistry, Czech Academy of Sciences, 5Institute of Animal Physiology and Genetics, Czech Academy of Sciences, 6Department of Cell Biology, Faculty of Science, Charles University, 7Institute of Experimental Medicine, Czech Academy of Sciences, 8Center for Eye Research, Department of Ophthalmology, Oslo University Hospital and Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, 9Moorfields Eye Hospitals UAE, 10Stem Cell Therapies in Neurodegenerative Diseases Lab, Research Center “Principe Felipe”, 11Eye Center Donaustadt, Department of Ophthalmology, Sigmund Freud University

The subretinal implantation of retinal pigmented epithelium (RPE) is one of the most promising approaches for the treatment of degenerative retinal diseases. However, the performance of preclinical studies on large-eye animal models remains challenging. This report presents guidelines for the subretinal transplantation of RPE on a cell carrier into minipigs.

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