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Optimization of a CRISPR/Cas9 genetic deletion system in macrophages to evaluate proteins important for the activation of NLRC4 inflammasome in macrophages

Grant number: 20/06177-3
Support type:Scholarships in Brazil - Master
Effective date (Start): October 01, 2020
Effective date (End): February 28, 2022
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal researcher:Dario Simões Zamboni
Grantee:Letícia de Sousa Lopes
Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:19/11342-6 - Mechanisms and consequences of the activation of cytoplasmic receptors by intracellular pathogens, AP.TEM

Abstract

Inflammasome is a multiproteic complex that assists the activation of various inflammatory caspases in response to environmental signals or the entry of pathogens into a cell. Soon after the recognition of the ligand, the inflammasome complex is formed and promote the cleavage of pro-IL-1b, pro-IL-18 and gasdermin D. The activation of the inflammasome leads to cytokine secretion and inflammatory cell death, also known as pyroptosis. Thus, the inflammasome plays a crucial role in the defense of the host against intracellular pathogens, such as Salmonella, Shigella, Legionella pneumophila and Pseudomonas aeruginosa, besides being related to the appearance and progression of hereditary or inflammatory diseases, such as the so-called MAS (Macrophage Activation Syndrome), neonatal-onset enterocolitis, and glioma. Despite being one of the most studied inflammasomes, some mechanisms involved in the NLRC4 pathway still need to be identified. By expanding knowledge about molecules involved in their activation and signaling, it will be possible to achieve a better understanding of inflammatory diseases related to it, as well as the role of the inflammasome in the control of intracellular pathogens. In this project, we intend to use the CRISPR-Cas9 technique to evaluate the role specific genes identified in a screening in the lab for proteins that interacts with NLRC4 in response to Legionella pneumophila infection. (AU)

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