Generation of iPSCs by reprogramming Gorilla gorilla fibroblasts

• Grant number: 25/11258-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Fabiana Fernandes Bressan
• Grantee: Kauã Weik Orneles de Freitas
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil

Abstract

The gorilla is the largest of the primates and, with the exception of the chimpanzee, is the closest relative to humans. Their genetic proximity allows for evolutionary and adaptive studies to be carried out, to understand the genes related to diseases, mechanisms of resistance to pathogens, the development of gene therapies, as well as the possibility for use in translational research, biomedical applications, and conservation of the species, which is classified as "Critically Endangered" by the IUCN. An efficient tool in these studies is the production of induced pluripotent stem cells (iPSCs), which can be generated via an integrative methodology, which results in the insertion of vectors into the DNA of the reprogrammed cell, rather than integrative, in which the vectors reach the cells through plasmids, making them free of transgenes. However, episomal reprogramming from fibroblasts in the western gorilla species has not yet been reported, due to the difficulty in obtaining biological material for the study of this species. In this proposal, the objective is to compare the generation of iPSCs generated through the reprogramming of fibroblasts previously isolated from Gorilla gorilla using the non-integrative reprogramming method, using episomal vectors (pMASTER 12 and pCXB EBNA1) and integrative, using a lentiviral vector with human transcription factors OSKM transcription factors (OCT4, SOX2, c-MYC, and KLF4). If colonies are present, they will be characterized based on morphology, gene expression, formation of embryoid bodies, alkaline phosphatase, and immunocytochemistry for pluripotency factors.