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

Grant number: 23/10975-0
Support Opportunities:Scholarships in Brazil - Support Program for Fixating Young Doctors
Effective date (Start): September 01, 2023
Effective date (End): August 31, 2025
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Acordo de Cooperação: CNPq
Principal Investigator:Ana Marisa Chudzinski-Tavassi
Grantee:Mariana Barbosa de Souza Rizzo
Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
Associated research grant:23/01532-8 - Neuroblastoma gene editing by CRISPR/Cas9 for target validation involved in pain and inflammation, AP.R

Abstract

INTRODUCTION: 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 antinociceptive 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 antinociceptive 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.

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