RT info:eu-repo/semantics/article T1 CRISPR/CasRx proof-of-concept for RNA degradation: A future tool against RNA viruses? A1 Pérez San José, Diana A1 Fuente García, Miguel Ángel de la A1 Serna Pérez, Julia A1 Simarro Grande, María A1 Eiros Bouza, José María A1 Sanz Muñoz, Iván K1 Biología K1 Virology K1 Infectious Diseases K1 Immunology K1 CRISPR/Cas system K1 Antiviral K1 Influenza virus K1 CRISPR/CasRx K1 2415 Biología Molecular AB Influenza viruses provide a great threat for the human population, causing highly contagious respiratory infections that can lead to serious clinical complications. There are a limited variety of influenza antivirals, and these antivirals are subjected to the constant emergence of resistances. Therefore, the development of new antiviral strategies to combat influenza viruses and other RNA viruses must be promoted. In this work, we design a proof-of-concept of a recently described CRISPR/Cas tool that has been proposed as a possible future RNA virus antiviral, named CRISPR/CasRx. For this, we verified the efficiency of the CasRx endonuclease in the degradation of the eGFP mRNA reporter gene and we established the best conditions for, and the efficient performance of, the CRISPR/CasRx system. The results were measured by fluorescence microscopy, flow cytometry, and qRT-PCR. The analyses demonstrated a reduction in fluorescence, regardless of the amount of eGFP reporter plasmid transfected. The analyses showed an 86–90% reduction in fluorescence by flow cytometry and a 51–80% reduction in mRNA expression by qRT-PCR. Our results demonstrate that the CasRx endonuclease is an efficient tool for eGFP mRNA knockdown. Therefore, subsequent experiments could be useful for the development of a new antiviral tool. PB MDPI YR 2021 FD 2021 LK https://uvadoc.uva.es/handle/10324/59136 UL https://uvadoc.uva.es/handle/10324/59136 LA eng NO Pharmaceuticals, 2022, vol. 15, n. 1, 32 NO Producción Científica DS UVaDOC RD 14-dic-2024