• crisprcas9
• nsd1
• nsd1a
• nsd1b
• sindrome di sotos
• zebrafish

NSD1 gene encodes for a protein with N-methyltransferase activity, which regulates gene transcription mainly through exterting histone H3 mono- and demethylation on lysine 36. NSD1 haploinsufficiency is the main cause of Sotos syndrome, an autosomal dominant disorder of neurodevelopment, characterized by abnormal growth and and altered behaviours. Two NSD1 orthologues are present in zebrafish, nsd1a and nsd1b, which show high homology to the human protein. The aim of this thesis is to develop a loss-of-function model for NSD1 orthologs using CRISPR/Cas9 gene editing technique and to create customised disease models by Base Editing. Crispants are generated by microinjecting guide RNA and Cas9 as ribonucleoprotein in zebrafish zygotes. Subsequent analyses include phenotypic, molecular and behavioural studies to assess the possible reseambling of traits of Sotos syndrome patients, including heart defects and aggressiveness. Proof of concept experiments support the use of Base Editing approachto create zebrafish diseases models with specific pathogenic NSD1 variants, paving the way for personalised medicine.