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HAVBRUK2-Stort program for havbruksforskning

Gene editing for elucidating gene function and refining genomic selection for CMS resistance in Atlantic salmon

Alternative title: Genredigering for å utforske genfunksjon og videreutvikle genomseleksjon for resistens mot CMS hos atlantisk laks

Awarded: NOK 10.0 mill.

Gene editing is a new technology that has potential to provide new opportunities for breeding to improve production efficiency and diseases resistance. Cardiomyopathy syndrome (CMS) is a disease that causes economic loss and affects the welfare of fish grown by the aquaculture industry in Norway. Selective breeding is currently used to mitigate the impact of the disease on the industry, but effective implementation of new technologies could potentially make selective breeding much more efficient. Previous studies have located areas in the genome that contain genes affecting CMS resistance. Use of state-of-the-art gene editing technologies for improvement of CMS resistance is likely to be highly beneficial for Atlantic salmon aquaculture. The project will identify potential genes and causative mutations affecting CMS resistance, and evaluate their function in laboratory cultured heart cells. The proposal will also develop and assess the effectiveness of breeding strategies employing gene editing for creating CMS resistant salmon, and design and assess the potential of a special scheme aiming to deliver the edited genes to salmon fingerling suppliers. The ethical, social and legal implications of developing CMS resistant salmon using gene editing will be investigated. Active responsible research and innovation processes for gene editing will be promoted with stakeholders. The project will create a knowledge base and example for the industry regarding disease management through state-of-the-art gene editing, this will greatly improve the bioeconomy and welfare of Atlantic salmon production. Whole genome sequencing and fine mapping Samples from three Mowi populations were selected based on prior knowledge on CMS related phenotypes and breeding values of the individuals. Whole genome sequencing was performed on samples from ~200 selected individuals (~100 per extreme phenotypes) and the sequence data were analyzed. A genomic region that is associated with CMS resistance is confirmed at chromosome 27 following fine mapping methodologies. PCR (polymerase chain reaction) based results have validates the identified variant. Pilot micro injection of salmon eggs CRISPR/Cas9 constructs were designed based on the results from the sequence analysis and prior knowledge of the chromosomal region. A total of six CRISPR/Cas9 constructs targeting three genes (two per gene) were designed. A pilot micro injection of salmon eggs was carried out to test efficiency of the CRISPR/Cas9 constructs before the main injection trial and validation process to confirm the success of CRISPR edit are being carried out through amplicon sequencing and in vitro cell culture. Engaging citizens in research and innovation: mutual learning workshop With the aim of better understanding citizens' attitudes and perception towards gene editing in salmon, a ?Science Espresso? format workshop was conducted. Participants with different geographical, political, and religious backgrounds, representing diverse ethical views and food-related interests, were recruited to participate in a workshop. Results showed that lay citizens could raise important questions, give opinions on the technology, research objectives, and approach. Researchers can better understand attitudes, barriers, and opportunities; new questions were raised on both sides, and dialogue was established. Ethics of CMS resistance salmon: ethical matrix workshop An ethical matrix is a conceptual tool designed to map ethical issues, facilitate ethical reflection and assist decision-makers in decisions about the ethical issues regarding existing or prospective technologies in the field of food and agriculture. During a workshop in the CMSEdit consortium, the members of the project were put in small groups to identify the ethical issues involved in applying CRISPR for achieving resistance to cardiomyopathy syndrome (CMS) amongst farmed salmon. The Ethical Matrix assists in analyzing ethical issues for technologies in food and agriculture. The benefits for CMS resistant salmon are to be found in a potential for less loss of life which again causes benefits for the entire value chain in fish farming as resources are saved. However, there are paradoxes here since more fish will experience suffering through treatments and farming practices. Furthermore, ethical issues are connected to the price of CMS resistant salmon as well as to how the intellectual property rights are handled. These issues relate to consumer choice, farmers ?access to eggs or smolt that are CMS resistant, and the possibility of introducing ecological alternatives. A foundational ethical issue is whether gene editing

Gene editing technologies are developing at a faster pace than ever and open new opportunities for breeding to improve production efficiency and diseases resistance. Cardiomyopathy syndrome (CMS) is an important disease for the aquaculture industry in Norway because of economic loss and welfare implications. To mitigate the impact of the disease selective breeding is practiced, however, it is challenged by veterinary restrictions and expensive challenge tests. Genomic studies for the trait have located quantitative trait loci (QTL) with strong signal on chromosome 27. The use of state-of-the-art gene editing technologies for improvement of CSM resistance is likely to be highly beneficial in this case. The project will identify potential genes and causative mutations affecting CMS resistance, and evaluates their function by in-vitro cardiomyocyte cell culture. Further, the proposal will develop breeding strategies that effectively employ gene editing to develop CMS resistant salmon, and a special scheme to deliver edited genes to multipliers tiers, which produces fingerlings that are grown for production, will be designed. The ethical, social and legal implications of developing CMS resistant salmon using gene editing will be determined. Dynamic RRI (responsible research and innovation) processes, in particular active stakeholder involvement, are used to promote an active attitude towards RRI in gene editing. The project will create a knowledge base and example for the industry regarding disease management through state-of-the-art gene editing, this will greatly improve the bioeconomy and welfare of Atlantic salmon production.

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HAVBRUK2-Stort program for havbruksforskning