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

鲑鱼慢性病毒病原体疫苗——干扰素减毒细胞系的产生

• 批准号: NE/P010946/1
• 负责人: Samuel Martin
• 金额: $ 27.29万
• 依托单位: University of Aberdeen
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NE%2FP010946%2F1
• 关键词: Vaccines; chronic; viral; pathogens; salmon

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

鲑鱼慢性病毒病原体疫苗-干扰素减毒细胞系的产生。Acroym：SalVacCell项目目标/主要目标。鲑鱼水产养殖业迫切需要生产针对病毒病原体的新疫苗。病毒是鲑鱼水产养殖业的主要经济损失之一。这直接反映了鲑鱼产业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，但目前这个项目将使用细胞系。经过 CRISPR/cas9 编辑，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，这是动物体内​​主要的抗病毒机制。这些细胞系还将用于解释与鲑鱼天然病毒抗性/易感性相关的基因的基本功能，这对育种计划具有直接重要性。我们已经获得了工业合作伙伴 Benchmark（疫苗公司）和 Landcatch。 （鱼类育种）。 目标 1. 通过敲除 IFN 功能来开发 IFN 缺陷的鱼细胞系 2. 评估 KO 细胞系以产生抗病毒反应以确认表型 3。解释鱼类病毒自然产生的抗性/易感性的细胞系 传统细胞系和新开发的细胞系之间病毒颗粒产量和病毒诊断周转时间的比较 关键挑战 首要挑战目的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。许多实验室已尝试敲除鲑鱼细胞中的基因，但迄今为止尚未实现。据我们所知，我们是第一个创建此类细胞系的人，因此我们无资金支持的前期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，并大大改善疾病管理的基础。遗传选择。合作伙伴使用的缺乏抗病毒反应的细胞系。 b. 可用于生产疫苗的高滴度病毒的能力。鲑鱼养殖项目持续时间：24 个月项目总成本：354622 英镑（100%）BBSRC/NERC 请求捐款 249,622 英镑(£199697.60 80%)MSS 的贡献：获得 65K 英镑 Benchmark 的贡献获得 20K 英镑 Landcatch 的贡献获得 20K 英镑

项目成果

Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system

使用慢病毒传递系统对鲑鱼细胞系进行高效 CRISPR/Cas9 基因组编辑

• DOI: http://dx.10.1101/734442
• 发表时间: 2019
• 作者: Gratacap R

IFN Signaling in Inflammation and Viral Infections: New Insights from Fish Models.

炎症和病毒感染中的干扰素信号传导：来自鱼类模型的新见解。

• DOI: http://dx.10.3390/v11030302
• 发表时间: 2019
• 期刊: Viruses
• 作者: Langevin C

Viral Resistance and IFN Signaling in STAT2 Knockout Fish Cells.

STAT2 敲除鱼细胞中的病毒抗性和 IFN 信号转导。

• DOI: http://dx.10.4049/jimmunol.1801376
• 发表时间: 2019
• 期刊: 1950)
• 作者: Dehler CE

Efficient CRISPR/Cas9 genome editing in a salmonid fish cell line using a lentivirus delivery system.

使用慢病毒传递系统对鲑鱼细胞系进行高效 CRISPR/Cas9 基因组编辑。

• DOI: http://dx.10.1186/s12896-020-00626-x
• 发表时间: 2020
• 期刊: BMC biotechnology
• 影响因子: 3.5
• 作者: Gratacap RL