Complexation design of cationized gelatin and molecular beacon to visualize intracellular mRNA

Autor: Jun-ichiro Jo, Sho Takehana, Yasuhiko Tabata, Yuki Murata
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Cytotoxicity
Toxicology
Pathology and Laboratory Medicine
Biochemistry
Gelatin
Green fluorescent protein
Mice
Fluorescence Microscopy
Animal Products
Nucleic Acids
Medicine and Health Sciences
Fluorescence microscope
Microscopy
Multidisciplinary
Chemistry
Light Microscopy
Glyceraldehyde-3-Phosphate Dehydrogenases
Agriculture
Fluorescence
Laboratory Equipment
Engineering and Technology
Medicine
Biological Cultures
Research Article
Cell Culturing Techniques
food.ingredient
Imaging Techniques
Science
Equipment
Research and Analysis Methods
Transfection
Gelatin Media
Cell Line
food
Molecular beacon
Cations
Fluorescence Imaging
Animals
RNA
Messenger
Molecular Biology Techniques
Molecular Biology
Oligonucleotide
Biology and Life Sciences
Mesenchymal Stem Cells
Culture Media
Cell Hybridization
Biophysics
Nucleic acid
Zdroj: PLoS ONE, Vol 16, Iss 1, p e0245899 (2021)
PLoS ONE
ISSN: 1932-6203
Popis: The objective of this study is to prepare cationized gelatin-molecular beacon (MB) complexes for the visualization of intracellular messenger RNA (mRNA). The complexes were prepared from cationized gelatins with different extents of cationization and different mixing ratios of MB to cationized gelatin. The apparent size of complexes was almost similar, while the zeta potential was different among the complexes. Irrespective of the preparation conditions, the complexes had a sequence specificity against the target oligonucleotides in hybridization. The cytotoxicity and the amount of complexes internalized into cells increased with an increase in the cationization extent and the concentration of cationized gelatin. After the incubation with complexes prepared from cationized gelatin with the highest extent of cationization and at mixing ratios of 10 and 20 pmole MB/μg cationized gelatin, a high fluorescent intensity was detected. On the other hand, the complex prepared with the mixing ratio at 20 pmole/μg did not show any cytotoxicity. The complex was the most effective to visualize the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA endogenously present. In addition, even for enhanced green fluorescent protein (EGFP) mRNA exogenously transfected, the complex permitted to effectively detect it as well. It is concluded that both the endogenous and exogenous mRNA can be visualized in living cells by use of cationized gelatin-MB complexes designed.
Databáze: OpenAIRE
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