Vaccines for chronic viral pathogens in salmon- generation of interferon attenuated cell lines

鲑鱼中慢性病毒病原体的疫苗-干扰素减毒细胞系的产生。Acroym：SalVacCellProject goal / key aims。在鲑鱼水产养殖业中，主要需要产生针对病毒病原体的新疫苗。病毒是鲑鱼产业的主要经济损失之一，这直接反映了缺乏高保护性疫苗。为了改进疫苗设计和测试，需要大量的病毒，但目前这是不可能的。该项目将使用经过CRISPR/cas9编辑的细胞系，使其高度允许难以生长的病毒生长。我们将敲除I型干扰素途径，这是动物中主要的抗病毒机制。具体来说，我们将靶向调节干扰素诱导的细胞对病毒感染反应的关键基因。这些细胞系还将用于解释鲑鱼中与天然病毒抗性/易感性相关的基因的潜在功能，这对育种计划具有直接重要性。我们已获得工业合作伙伴Benchmark（疫苗公司）和Landcatch（鱼类育种）。联合体协议将明确定义IP的角色和保密性。通过干扰素功能敲除开发干扰素缺陷型鱼细胞系2。评估KO细胞系以产生抗病毒应答，从而确认表型3。使用细胞系来解释对鱼类病毒的自然发生的抗性/易感性4。比较传统细胞系和新开发细胞系之间的病毒颗粒生产产量和病毒诊断转换时间关键挑战首要挑战是能够为疫苗公司生产高滴度的病毒，以便能够改进对经济上重要的慢性病毒病原体的设计和保护。许多实验室试图敲除鲑鱼细胞中的基因，但迄今为止尚未实现。据我们所知，我们是第一个创建这种细胞系的人，因此我们没有资金的初步工作已经克服了一个主要障碍。该项目的挑战是充分利用我们的细胞系技术进行疾病管理，并大大改善遗传选择的基础。附件-供伙伴使用a。缺乏抗病毒应答的工程化细胞系。可用于生产疫苗用高滴度病毒的细胞系c。在工业环境中扩大病毒生产的能力。提高鲑鱼育种抗病性选择的精度项目持续时间：24个月项目总成本：354622英镑（100%）BBSRC/NERC要求的捐款249，622英镑（199697.60英镑80%）MSS的捐款：获得65，000英镑Benchmark的捐款获得2万英镑Landcatch的捐款获得2万英镑

Vaccines for chronic viral pathogens in salmon- generation of interferon attenuated cell lines.Acroym: SalVacCellProject goal / key aims.There is a major requirement to generate new vaccines for viral pathogens in the salmon aquaculture industry. Viruses represent one of the major economic losses to the salmon industry, which is a direct reflection of the lack of highly protective vaccines. In order to improve vaccine design and testing high quantities of viruses are required, but at present this is not possible. This project will use cell lines that have been edited by CRISPR/cas9 to be highly permissive for growth of difficult to grow viruses. We will knockout the type I interferon pathway which is the major antiviral mechanism in animals. Specifically we will target key genes that regulate interferon induced cellular responses to viral infection. These cell lines will additionally be used to explain the underlying function of genes associated with natural viral resistance / susceptibility in salmon that is of direct importance to breeding programs.We have secured Industrial partners Benchmark (vaccine company) and Landcatch (fish breeding). Consortium agreements will clearly define roles and confidentiality of IP.OBJECTIVES1. Development of IFN-deficient fish cell lines by knock-out of IFN function2. Evaluation KO cell lines to mount an antiviral response to confirm phenotype3. Use the cell lines to explain naturally occurring resistance / susceptibility to fish viruses4. Comparison of yield for viral particle production and viral diagnostic turn-over time between traditional cell lines and newly developed cell lines KEY CHALLENGESThe overarching challenge is to be able to produce high titres of virus for vaccine companies to be able to improve design and protection to economically important chronic viral pathogens. Many labs have attempted to knock down genes in salmonid cells, but to date this has not been achieved. To our knowledge we are the first to create such cell lines and as such our unfunded preliminary work has already overcome a major hurdle. The challenge of this project is to fully exploit our cell line technology for both disease management and also to greatly improve the basis of genetic selection. Deliverables- to be used by partnersa. Engineered cell lines that are deficient in antiviral responses.b. Cells lines that can be used to produce high titre of viruses for vaccinesc. Capacity to upscale viral production in industrial environment.d. Improved precision of selection for disease resistance in salmon breedingProject duration: 24 monthsTotal project cost: £354622 Contribution requested from (100%) BBSRC/NERC £249,622 (£199697.60 80%)Contribution from MSS: secured £65KContribution from Benchmark secured £20K Contribution from Landcatch secured £20K