Cloning of cone-specific inducible expression using CRISPR Cre-Lox system

• Grant number: 25/11456-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: February 01, 2026
• End date: January 31, 2027
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Alexandre Hiroaki Kihara
• Grantee: Théo Henrique de Lima Vasconcellos
• Supervisor: David e Cobrinik
• Host Institution: Centro de Matemática, Computação e Cognição (CMCC). Universidade Federal do ABC (UFABC). Santo André , SP, Brazil
• Institution abroad: University of Southern California (USC), United States
• Associated to the scholarship: 24/00828-3 - Utilizing flavonoids for the treatment of autosomal recessive retinitis pigmentosa: a nutraceutical approach, BP.DR

Abstract

Retinal organoids (ROs) provide a remarkable platform for studying retinal development and diseases. These self-organized structures resemble native retinal tissue, encompassing various cell types and functioning photoreceptors. ROs enable the modeling of retinal diseases using patient-derived cells, induced pluripotent stem cells (iPSCs), offering insights into disease mechanisms and potential therapeutic strategies. In this project, we aim to delve into advanced gene editing techniques combining CRISPR/Cas9 and Cre-Lox systems. CRISPR/Cas9 offers precise DNA modification, while the Cre-Lox system facilitates controlled gene activation. The focus is on developing an inducible Cre-Lox system in cone photoreceptors, enabling specific gene activation using tamoxifen. The primary objective of this project is to integrate the estrogen receptor T2 (ERT2)-Cre-ERT2 cassette into the G-protein subunit alpha transducin 2 (GNAT2) gene location and the CAG-LSL-GOI-T2A-Maroon sequence into the AAVS1 gene location of iPSCs. Subsequently, the focus will be on differentiating these iPSC lines into retinal organoids. This approach holds potential for disease modeling and therapeutic screening. In summary, the project provides a deep understanding of retinal morphology, cellular interactions, and gene editing techniques. It highlights the potential of retinal organoids and inducible genetic systems for advancing our understanding of retinal development, disease mechanisms, and potential treatments.