Eliminação do gene RASSF9 na linhagem de melanoma B16F0 através de CRISPR/Cas9

• Grant number: 22/05193-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2022
• End date: June 30, 2023
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: João Gustavo Pessini Amarante Mendes
• Grantee: Julia Souza e Costa
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Melanoma is a form of cancer caused by the transformation of melanocytes. Importantly, melanoma is the most lethal among all skin cancers. Melanoma can be divided into four groups depending on the type of mutation present in the melanoma cells. The more frequent alterations were mapped in B-raf and ras genes. The Ras protein may initiate different signaling cascades, thereby regulating processes such as proliferation, differentiation, morphology, and apoptosis. The main effectors of Ras related to cell death are members of the RASSF family (Ras-association Family). RASSF9 has been implicated in keratinocyte differentiation and dermal homeostasis and, interestingly, its expression can be induced by sun exposure. To study the role of RASSF9 in melanomas, we will use the CRISPR/Cas9 system to eliminate the expression of this gene in the murine lineage B16F0. After selecting stable transfectants, we will evaluate the new melanoma strain (B16F0.R9KO) regarding in vitro and in vivo tumor growth, as well as resistance to chemotherapy. Finally, the B16F0.R9KO strain will serve as a tool for further studies on the role of RASSF9 on intracellular biochemical pathways and other aspects of tumor biology.(AU)