Improving Magnetotransfection through FACS cell enrichment and its application on fish cells

• Grant number: 24/22020-8
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 28, 2025
• End date: August 27, 2025
• Field of knowledge: Agronomical Sciences - Fishery Resources and Fishery Engineering - Aquaculture
• Principal Investigator: Maeli Dal Pai
• Grantee: Erika Stefani Perez
• Supervisor: Daniel Garcia de La Serrana Castillo
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Universitat de Barcelona (UB), Spain
• Associated to the scholarship: 21/13905-8 - Transcriptomic and functional analysis as strategies for understanding the determinate and indeterminate muscle growth patterns of fish, BP.DR

Abstract

Muscle growth in fish occurs by hypertrophy and hyperplasia. The hyperplasia can be classified in two types: mosaic and stratified. Fish with mosaic hyperplasia continuous throughout life have indeterminate growth, on the other hand, fish with mosaic hyperplasia more restricted to the first days after hatching, have determined growth. Numerous studies carried out over the years have contributed to increase knowledge about muscle growth, however, the indeterminate growth pattern is not fully understood. In this sense, functional analyses can represent efficient tools to prove the genes function selected in silico and, involved in indeterminate growth. These experiments involving the manipulation of gene expression are relevant in the aquaculture scenario, given the constant demand for increased and improved production. In this context, this project aims to manipulate the transcription of wasla and/or waslb genes, predicted to be involved in skeletal muscle growth, in the European seabass (Dicentrarchus labrax), an indeterminate growth species. For this proposal, we will use primary cell culture of myoblasts. These cells will be magnetotransfected using the protocol pre-established in the previous BEPE project, and enriched through FACS to have a 100% transfected cells. Considering the robust methodology that will be used, it is expected that the knowledge generated by this project can complement, in a very significant way, the results that are being obtained in the doctoral project funded by FAPESP (2021/13905-8). In addition, it can contribute with the development of innovation projects that are of great interest to the productive sector. Thinking in the technological learning, the magnetotransfection and FACS could be applied in Brazil using other species of fish