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SynBac: Synthetic Baculovirus Genome for Next-generation Drug Discovery

SynBac：用于下一代药物发现的合成杆状病毒基因组

基本信息

批准号：
1864756
负责人：
金额：
--
依托单位：
University of Bristol
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2016
资助国家：
英国
起止时间：
2016 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-1864756
关键词：
SynBac Synthetic Baculovirus Genome Next

项目摘要

Baculovirus is a highly efficient delivery system for recombinant genes into eukaryotic cells, with great impact on the production of eukaryotic proteins, including high-value drug targets. Vaccines against cervicular cancer and others are produced by this method. More recently, baculovirus has emerged as a versatile tool for gene therapy. We contributed to the field the award-winning MultiBac technology for multiprotein complex research. These applications, at the forefront of modern biology, rely on a large baculovirus genome (130 kb) derived from AcMNPV. This genome has been intensively researched, mainly by entymologists. Genes essential for propagation in nature and in cell culture were delineated and DNA elements which impede applications in the laboratory. Genetic alterations of the wild-type viral genome have been performed, by classical knock-out technologies, to improve gene insertion, delivery and protein production properties. Such alterations require excessive effort by specialists. Therefore, it is currently not possible to fully exploit the vast potential of the baculovirus system. In the present project, we boldly propose to fully reverse the current approach. We will design in silico and construct in vitro new, fully synthetic customized baculovirus genomes which will be, for the first time, in a streamlined, highly versatile format for multigene transfer and the production of high-value, next generation recombinant protein targets for drug discovery. We will apply state-of-the-art genome editing tools, notably CRISPR-Cas9, to inform our approach by systematically disrupting genes and non-coding regions including gene regulatory elements. We further aim to address the "scale-up problem" which currently impedes pharma-scale biologics production by baculoviral systems. As proof-of-concept, we already created a partly synthetic hybrid genome by replacing a large part (20 kb) of wild-type with designer DNA. Rigorous testing of this prototype compellingly validated our approach.The CASE fellow will:(1) utilize computational biology, comparative bioinformatics and data mining to create blueprints for optimized minimal baculovirus genomes. (2) Synthesize designer genomes in large fragments, and use advanced recombination technologies to assemble these into functional genomes (AstraZeneca platform).(3) Exploit cutting-edge CRISPR-Cas9 tool to edit genes and regulatory DNA elements in the baculoviral genome in a parallelized fashion (AstraZeneca platform) and implement this information in the synthetic design. (4) Address the "scale-up problem" by reconfiguring the very late phase of baculoviral life cycle ("hyperburst management"). (5) Rigorously validate novel designer genomes experimentally.These will be the first fully synthetic baculoviral genomes, with the potential to transform academic and industrial R&D applications.

杆状病毒是一种将重组基因高效导入真核细胞的载体系统，对真核蛋白的产生有很大影响，包括高价值的药物靶标。宫颈癌和其他癌症的疫苗就是用这种方法生产的。最近，杆状病毒已成为基因治疗的通用工具。我们为多蛋白复合物研究领域贡献了屡获殊荣的MultiBac技术。这些应用，在现代生物学的最前沿，依赖于一个大的杆状病毒基因组（130 kb）来自AcMNPV。该基因组已得到深入研究，主要由昆虫学家进行。描述了在自然界和细胞培养中繁殖所必需的基因，以及阻碍实验室应用的DNA元件。已经通过经典的敲除技术对野生型病毒基因组进行遗传改变，以改善基因插入、递送和蛋白质产生特性。这种改变需要专家的过度努力。因此，目前不可能充分利用杆状病毒系统的巨大潜力。在本项目中，我们大胆地提出完全扭转目前的做法。我们将通过计算机模拟设计并在体外构建新的、完全合成的定制杆状病毒基因组，这些基因组将首次以简化的、高度通用的形式用于多基因转移和生产高价值的下一代重组蛋白靶点。药物发现。我们将应用最先进的基因组编辑工具，特别是CRISPR-Cas9，通过系统地破坏基因和非编码区（包括基因调控元件）来为我们的方法提供信息。我们进一步的目标是解决目前阻碍杆状病毒系统的制药规模的生物制剂生产的“规模扩大问题”。作为概念验证，我们已经通过用设计者DNA替换野生型的大部分（20 kb）来创建部分合成的杂交基因组。该原型的严格测试令人信服地验证了我们的方法。CASE研究员将：（1）利用计算生物学，比较生物信息学和数据挖掘来创建优化的最小杆状病毒基因组的蓝图。(2)以大片段合成设计基因组，并使用先进的重组技术将其组装成功能基因组（阿斯利康平台）。(3)利用尖端的CRISPR-Cas9工具以并行方式编辑杆状病毒基因组中的基因和调控DNA元件（阿斯利康平台），并在合成设计中实施这些信息。(4)通过重组杆状病毒生命周期的最后阶段（“超爆发管理”）解决“规模扩大问题”。(5)通过实验严格验证新型设计基因组。这将是第一个完全合成的杆状病毒基因组，具有改变学术和工业研发应用的潜力。