RT info:eu-repo/semantics/doctoralThesis
T1 Translational development of an antiviral treatment against respiratory RNA viruses based on CRISPR-CasRx technology
A1 Pérez San José, Diana
A2 Universidad de Valladolid. Escuela de Doctorado
K1 Microbiología
K1 influenza viruses
K1 Influenza virus
K1 CRISPR-Cas13d
K1 SARS-CoV-2
K1 3109.05 Microbiología
AB Influenza viruses are a common cause of respiratory tract infections, causing epidemics each year. However, as historical events have demonstrated, they also carry the potential to cause global pandemics. The impact of these viruses on global health and its socioeconomic effects are significant. The best prophylactic strategy against it is vaccination, although the low percentage of vaccination in the population affects its effectiveness, which is also decreased due to sudden antigenic drifts in the viral glycoproteins. In severe cases of infection, the use of antiviral drugs is recommended, to lessen the symptoms and improve the prognosis of the disease. However, there is currently a lack of effective antiviral drugs, as all available options are susceptible to the development of resistances. For this reason, there is a need to develop new antivirals that are easily adaptable to specific mutations of epidemic viruses and that can be produced quickly and cheaply so that they can be adapted to the sudden appearance of mutations.These novel antivirals could also prove valuable in pandemic scenarios, given their potential for rapid design and production. The recent COVID-19 pandemic exemplifies the urgent need for such adaptable therapeutic strategies. Overall, there remains a pressing need to expand and improve the current arsenal of approved antivirals targeting respiratory viruses in the human population.In this study, we propose the CRISPR-CasRx system as a novel antiviral mechanism. Due to its programmability, CasRx can be readily adapted to target the genome sequences of emerging RNA viruses. Promising advances in delivery methods, particularly using adeno-associated viral vectors, further support its potential for therapeutic use. We evaluated multiple guide RNAs targeting the influenza virus hemagglutinin and neuraminidase, as well as the SARS-CoV-2 PLpro. Our results highlight the critical influence of guide RNA design on CasRx activity, emphasizing the importance of careful target sequence selection to achieve optimal RNA cleavage. Most of the guides tested resulted in statistically significant reductions in mRNA and protein levels of the target sequences. Although further antiviral assays are needed, our data demonstrate promising results in the reduction of the viral load in influenza in vitro models. And a clear reduction in both mRNA and protein expression in SARS-CoV-2 antiviral assays, supporting their potential as antiviral tools.Overall, the findings presented in this thesis highlight the potential of CRISPR-CasRx systems as a novel antiviral strategy against respiratory RNA viruses. While further studies are needed to confirm their impact on viral progeny and to optimize delivery systems, particularly for in vivo applications, these results lay important groundwork for the development of adaptable and effective RNA-targeting antiviral therapies.
YR 2024
FD 2024
LK https://uvadoc.uva.es/handle/10324/80032
UL https://uvadoc.uva.es/handle/10324/80032
LA eng
NO Escuela de Doctorado
DS UVaDOC
RD 29-nov-2025