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Czech Academy of Sciences

4 ARTICLES PUBLISHED IN JoVE

Neuroscience

Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice

Takahiro Tadokoro¹, Atsushi Miyanohara¹, Michael Navarro¹, Kota Kamizato¹, Stefan Juhas², Jana Juhasova², Silvia Marsala¹, Oleksandr Platoshyn¹, Erik Curtis³, Brandon Gabel³, Joseph Ciacci³, Nada Lukacova⁴, Katarina Bimbova⁴, 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.

Biology

Practical Use of RNA Interference: Oral Delivery of Double-stranded RNA in Liposome Carriers for Cockroaches

Jia-Hsin Huang¹, Yun Liu¹, Yu-Hsien Lin^1,2,3, Xavier Belles⁴, How-Jing Lee¹

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.

Biochemistry

Spectrophotometric Determination of Phycobiliprotein Content in Cyanobacterium Synechocystis

Tomáš Zavřel¹, Dominik Chmelík^1,2, Maria A. Sinetova³, Jan Červený¹

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.

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 Studenovska⁴, David Rais⁴, Štěpán Popelka⁴, Lucie Tichotová^5,6, Yaroslav Nemesh^5,6, Anastasiia Kolesnikova^5,6, Ruslan Nyshchuk^5,6, Anna Brymová^5,6, Zdeňka Ellederová⁵, Jana Čížková⁵, Jana Juhásová⁵, Štefan Juhás⁵, Pavla Jendelová⁷, Richárd Nagymihály⁸, Igor Kozak⁹, Slaven Erceg¹⁰, Susanne Binder¹¹, Brigitte Müller¹, Knut Stieger¹, Jan Motlik⁵, 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.