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

鲑鱼慢性病毒病原体疫苗--产生干扰素使细胞系变弱。Acroym：SalVacCellProject目标/主要目标。在鲑鱼水产养殖业中，生产新的病毒病原体疫苗是一个主要需求。病毒是三文鱼养殖业的主要经济损失之一，这直接反映了缺乏高度保护性的疫苗。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。该项目将使用已经由CRISPR/Cas9编辑的细胞系，以高度允许难以生长的病毒的生长。我们将敲除I型干扰素途径，这是动物体内主要的抗病毒机制。具体地说，我们将针对调节干扰素诱导的细胞对病毒感染的反应的关键基因。这些细胞系还将用于解释与鲑鱼天然病毒抗性/敏感性相关的基因的潜在功能，这对育种计划直接重要。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知识产权的角色和机密性。通过敲除干扰素功能建立干扰素缺陷型鱼类细胞系2。评估KO细胞系的抗病毒反应以确认表型3。用这些细胞系来解释自然产生的对鱼类病毒的抗性/敏感性4。传统细胞系和新开发的细胞系之间的病毒颗粒产量和病毒诊断周转时间的比较关键的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。许多实验室试图破坏鲑鱼细胞中的基因，但到目前为止还没有实现。据我们所知，我们是第一个建立这种细胞系的人，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，并极大地改善基因选择的基础。交付成果-由Partnersa使用。缺乏抗病毒反应的基因工程细胞系。可用于生产用于疫苗接种的高滴度病毒的细胞系。在工业环境中扩大病毒生产的能力。提高了鲑鱼养殖抗病选择的精确度项目持续时间：24个月项目总成本：GB 354622要求(100%)BBSRC/NERC GB 249,622(GB 199697.60 80%)来自MS的贡献：保证GB 65K来自基准的贡献保证GB 20K来自LandCatch的贡献保证GB 20K

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