CRISPR/Cas9 Nickase as Strategy for Correcting the Pathogenic Variant c.5047C>T Responsible for Recessive Dystrophic Epidermolysis Bullosa

• Grant number: 25/05454-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Priscila Keiko Matsumoto Martin
• Grantee: Laurent Ketlen Leão Viana
• Host Institution: Instituto Israelita de Ensino e Pesquisa Albert Einstein (IIEPAE). Sociedade Beneficente Israelita Brasileira Albert Einstein (SBIBAE). São Paulo , SP, Brazil

Abstract

Epidermolysis Bullosa (EB) is a rare genetic dermatological disorder characterized primarily by the formation of blisters and erosions resulting from minor friction on fragile skin, which manifest shortly after birth. To date, there is no cure for this condition. EB is classified into four classical subtypes: EB Simplex (EBS), Junctional EB (JEB), Dystrophic EB (DEB), and Kindler EB (KEB), each of which exhibits three degrees of phenotypic severity. Recessive Dystrophic Epidermolysis Bullosa (RDEB) is one of the most severe subtypes, caused by mutations in the COL7A1 gene, which encodes type VII collagen, the primary protein of anchoring fibrils and a critical structural component for dermal-epidermal adhesion. In Brazil, the c.5047C>T mutation is one of the most prevalent and arises from the formation of a premature stop codon, leading to the interruption of gene transcription and resulting in deficient production of type VII collagen. Currently, several studies exploring gene therapy, CRISPR/Cas9, and its derivatives are being investigated as potential therapeutic approaches for RDEB. The primary objective of this project is to optimize the CRISPR/Cas9n nickase editing tool for the correction of the pathogenic variant c.5047C>T. This tool holds significant promise, particularly with improvements in homologous recombination through the design of 5' and 3' overhang oligonucleotide strategies. Furthermore, it is essential to develop enhanced tools that can be adapted for other EB-causing variants, including deletions and insertions in the COL7A1 gene.