PETRI-fish project

PETRI-fish: CRISPR-screening to reduce challenge testing in aquaculture.   

Duration: April 2024 – September 2026

Responsible: Matthew Kent

Funding source: FHF

Project number: 901918

Awarded: 7.2MNOK

Main objective:

PERTI-FISH will address the widespread use of live fish testing and the intrinsic limitations of using production fish by offering a scalable, in-vitro approach that is not limited to pre-existing genetic variation. We will build upon our existing experience performing CRISPR-screening in livestock and apply and adapt this to salmonids including Atlantic salmon to answer the following questions:

• Which salmonid cells exist or can be created that can be used for in vitro testing?
• How can cells be mass transduced efficiently?
• What are the ideal parameters when developing CRISPR-screening libraries?
• Which genes or pathways are involved in ISAV resistance and susceptibility in salmonids?

We will go beyond delivering protocols and use the technology to investigate genes underlying resistance and susceptibility to ISAV, a tractable and highly relevant disease. Importantly, we assert that the principles and approaches we develop can be transferred to other diseases further reducing the need for live animal testing.

Summary:

The PETRI-FISH project responds to the call for research projects by The Norwegian Seafood Research Fund (FHF) on Utvikling av 3R forsøksmodeller og metodikk i havbruksnæringen. The project will focus on developing methods and models that have real potential to reduce the number of fish used in experimental testing by replacing live disease challenge testing with novel in vitro testing approaches.

CRISPR-screening is an extremely powerful tool for unbiased discovery of phenotype relevant genes and is already well established in human and mouse, with screens to identify essential genes, discover genes related to cancer development, and discover novel host-pathogen interactions for pathogens including Sars-CoV-2, dengue and zika virus, norovirus, and HIV to name a few; the approach is now transferring into non-model species.

Infectious Salmon Anaemia (ISA) caused by the ISA virus (ISAV) is among the most important diseases threatening the industry. Unfortunately, there is no fully effective treatment or vaccine, therefore genomics and breeding are explored as efficient alternatives or supplements for disease prevention.

Genome wide association studies (GWAS) indicate a low to moderate genetic heritability for ISA resistance, making it an attractive target for genetic improvement, however studies also reveal multiple genomic regions with small effects and despite evidence for some underlying genes, the functional basis for ISA resistance remains unknown.

The primary goal of this work is to apply CRISPR-screening to reduce live fish testing and to identify genes and pathways involved in ISAV disease resistance and susceptibility in Atlantic salmon.

Researchers involved: Assoc. prof. Matthew Kent, Dr. Thomas Harvey (co-PI), Prof. Sigbjørn Lien, Dr. Victor Boyartchuk, Dr. Tomasz Podgorniak, Prabin Sharma Humagain, Prof. Espen Rimstad, Dr. Jacob Torgersen.