CRISPR/Cas9 tool to knockout cell death pathway genes in neuronal cells and its impact under Zika virus infection

• Grant number: 19/09795-2
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Effective date (Start): June 01, 2019
• Effective date (End): May 31, 2020
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Ricardo Weinlich
• Grantee: Rafaela da Rosa Ribeiro
• Supervisor: Vania Broccoli
• Host Institution: Instituto Israelita de Ensino e Pesquisa Albert Einstein (IIEPAE). Sociedade Beneficente Israelita Brasileira Albert Einstein (SBIBAE). São Paulo , SP, Brazil
• Research place: Istituto di Ricovero e Cura a Carattere Scientifico S. de Bellis (IRCCS), Italy
• Associated to the scholarship: 17/03603-9 - The role of cell death pathways during the ZIKA virus infection, BP.PD

Abstract

Several pathways can trigger different kinds of cell death, being apoptosis, pyroptosis and necroptosis the most studied ones. The first two kinds involve caspases activation, while necroptosis is caspase independent and depends on the activation of a complex that contains RIPK3 and MLKL. During viral infection, the host cell triggers its own death trying to impair the virus replication niche. Nonetheless, coevolution of hosts and viruses gave viruses ways to block the cell death pathways. For example, herpes virus and poxvirus can block apoptosis and cytomegalovirus can block apoptosis and necroptosis. In 2015, ZIKA virus was brought to the global attention due its association with nervous system disorders, especially with microcephaly. It has been reported that ZIKA virus activates caspase-3, a central protein in the apoptosis pathway in human neuroprogenitor cells (NPC), but still is not well characterized how this caspase is activated. Moreover, little is known about the induction of other cell death pathways by ZIKA infection, especially regarding their importance to the disease phenotype. In this project, we intend to evaluate how the different cell death pathways impact in the phenotype of NPC infected with ZIKA virus. We will achieve that aim by knocking-out the neural stem cells, by CRISPR/Cas9, key genes in the different cell death pathways, such as FADD (central to the extrinsic apoptotic pathway), caspase-9 (intrinsic apoptotic pathway activator), caspase-1/11 (pyroptosis executors), RIPK3 and MLKL (necroptosis executors) and evaluating their differential susceptibility to the ZIKA infection. (AU)