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Neuroblastoma gene editing by CRISPR/Cas9 for target validation involved in pain and inflammation

Grant number: 23/01532-8
Support Opportunities:Regular Research Grants
Duration: July 01, 2023 - June 30, 2025
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Convênio/Acordo: CNPq
Principal Investigator:Ana Marisa Chudzinski-Tavassi
Grantee:Ana Marisa Chudzinski-Tavassi
Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Associated research grant:20/13139-0 - Centre of Excellence in New Target Discovery, AP.PCPE

Abstract

Pain is a worldwide problem. The identification and validation of targets affecting pain are critical for the development of more effective and safe interventions. Venoms are rich in bioactive compounds and can be valuable tools to explore new molecular targets. For pain studies, we characterized a model to evaluate the anti-nociceptive effect of compounds in human sensory-like neurons derived from SH-SY5Y cells, using a glycated extracellular matrix (ECM-GC) as stimulus. Venom derived peptides E8, D3, P4 and C-ambly induce anti-nociceptive phenotype in neurons by decreasing substance P and increasing ²-endorphin release. An analysis from chemoproteomics and bioinformatics led us to identify possible specific targets of peptides that participate in the downregulation of pain and inflammation, and these promising targets will be validated. OBJECTIVES: To validate therapeutic targets involved in pain and inflammation through gene knockout. METHODS: The KO of targets will be performed in the cell line SH-SY5Y using CRISPR/Cas9. The functional impact of deleted genes will be investigated using molecular and cellular biology approaches. EXPECTED RESULTS: We expect to observe how the targets contribute to the pain and inflammatory response of cells and how they are relevant for the action of the antinociceptive/anti-inflammatory peptides, which will potentially reveal a new specific target peptide to be used in inflammation and pain control. (AU)

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