Assessment of the impact of editing the sp7 and runx2a genes by CRISPR/Cas9 on the microRNA expression profile and the regenerative potential of zebrafish

• Grant number: 24/07320-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2024
• End date: July 31, 2025
• Field of knowledge: Biological Sciences - Genetics - Animal Genetics
• Principal Investigator: Danillo Pinhal
• Grantee: Maria Clara Tamascia de Castro
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

This research project aims to investigate the effects of editing the sp7 and runx2a genes on the epimorphic regeneration capacity of the caudal fin in zebrafish (Danio rerio), a valuable study model due to its unique ability to regenerate various types of tissues. Specific objectives include analyzing the expression profiles of edited genes (sp7 and runx2a) and regulatory microRNAs (miR-203a-3p, let-7c-3p, and miR-17-a1) in heterozygous mutant fish using real-time quantitative PCR (qPCR). Additionally, caudal fin regeneration will be compared between mutant and wild-type fish post-amputation, and the results will be correlated with morphometric analyses to understand the impact of genetic editing on regenerative capacity. This study is motivated by the importance of the sp7 and runx2a genes in the embryonic development of teleost fish, emphasizing the need to fill knowledge gaps regarding their roles in epimorphic regeneration. By better understanding how these genes and microRNAs influence regeneration, this project may provide valuable insights into tissue repair mechanisms in vertebrates, with potential future applications in regenerative medicine.