Non-Structural Protein Expression from West Nile Virus: Molecular detection of West Nile virus (WNV) and other arbovirus em msquitoes collected in Amazon region.

• Grant number: 24/21254-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2025
• End date: December 31, 2025
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Edison Luiz Durigon
• Grantee: Thais Pailo de Carvalho
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 23/07746-0 - Drug discovery against human viral infectious diseases, AP.TEM

Abstract

Brazil is one of the countries strongly haunted by parasitic diseases caused by viralpathogens. These infections severely affect public health and welfare in a wide part ofthe tropical world. Existing drug therapies are based on old and often very toxic activesubstances and reports on developing resistance of pathogens are increasing. Thesearch for new chemical entities with anti-pathogenetic activity and new mechanisms ofaction is thus a highly urgent task. Flaviviruses are important human pathogens andinclude dengue virus (DENV), West Nile virus (WNV), Japanese encephalitis virus (JEV),Yellow fever virus (YFV) and Zika virus (ZIKV) which are transmitted by mosquitoes andcause viral haemorrhagic fever and encephalitis in humans (Petersen et al., 2013,Flórez-Álvarez et al., 2022). Despite the significant impact of flavivirus infection onhuman health, vaccines are available for only a limited number of flaviviruses such asYFV, and antiviral therapies are not available for any flaviviruses. All flaviviruses carry apositive-sense single-stranded RNA genome, which encodes ten proteins; threestructural proteins form the virus shell, and seven non-structural (NS) proteins areinvolved in replication of the viral genome. While all NS proteins (NS1, NS2A, NS2B,NS3, NS4A, NS4B, and NS5) are part of a functional membrane-bound replicationcomplex, enzymatic activities required for flaviviral replication reside in only two NSproteins, NS3 and NS5. NS3 functions as a protease, helicase, and triphosphatase, andNS5 as a capping enzyme, methyltransferase, and RNA-dependent RNA polymerase.Within this doctoral project focus in WNV and other arbovirus isolated from Amazon region that will be given on the non-structural protein of WNV in terms of gene cloning, recombinant expression and preparation for proteincrystallisation.