Purpose of the STSM: The main purpose of this STSM was for the applicant to get familiar with the microinjection protocol in zebrafish to promote F0 knockout for a single gene. This is a crucial step for the overall success of a CRISPR-Cas9-based targeted gene knockout in this animal. By seeing it performed by experienced users, and being able to attempt it in a lab where the whole protocol is well-optimized, the chances for the applicant to latter implement that same method with success will greatly improve.
That is particularly relevant within the scope of her ongoing project “Neurological disease modelling for Sanfilippo: a key step towards understanding and treating a rare genetic disorder”(EXPL/BTM-SAL/0659/2021), which aims to create knockout zebrafish models for Sanfilippo to be used in the research for new RNA-based therapies. Thus, it was extremely important for the applicant to learn and perform this technology and then bring the knowledge and transfer it to other colleagues from Sandra Alves’ lab on Lysosomal Storage Disorders (Research and Development Unit, Human Genetics Department from the National Institute of Health Dr. Ricardo Jorge, in Portugal).

Grantee name: Maria Francisca Coutinho

STSM start and end date: 02/10/2022 to 06/10/2022

Title: A simple and effective F0 knockout method to generate zebrafish knock-out models for MPS III
Host lab: Prof. Stephen Wilson, University College London (UCL) Department of Cell and Developmental Biology (London, GB)