A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus.
A CRISPR/Cas9 and Cre/Lox system-based express vaccine development strategy against re-emerging Pseudorabies virus.
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DOI：
10.1038/srep19176
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发表时间：
2016-01-18
影响因子：
4.6
通讯作者：
Cao G
中科院分区：
文献类型：
--
作者：
Liang X;Sun L;Yu T;Pan Y;Wang D;Hu X;Fu Z;He Q;Cao G
病毒迅速进化以逃避疫苗诱导的免疫力，对有效的疫苗开发生物技术提出了迫切的需求。在这里，我们提出了一种基于CRISPR/Cas9和Cre/Lox系统的表达疫苗开发策略，以对抗最近在中国爆发的毁灭性猪伪狂犬病疫情的重新出现的伪狂犬病病毒。通过CRISPR/Cas9系统，新分离菌株的毒力基因同时被标记基因取代，随后使用Cre/Lox系统切除疫苗安全性问题。值得注意的是，应用单细胞FACS技术以进一步促进病毒纯化效率。这些先进技术的结合大大加速了疫苗的开发。最后，疫苗接种和攻毒实验证明了该候选疫苗在猪中的保护效果和控制当前伪狂犬病爆发的希望。据我们所知，这是第一个基于基因编辑技术的成功疫苗开发，表明这些技术从实验室到工业的飞跃。这可能为未来快速抗病毒疫苗的开发铺平道路。
Virus evolves rapidly to escape vaccine-induced immunity, posing a desperate demand for efficient vaccine development biotechnologies. Here we present an express vaccine development strategy based on CRISPR/Cas9 and Cre/Lox system against re-emerging Pseudorabies virus, which caused the recent devastating swine pseudorabies outbreak in China. By CRISPR/Cas9 system, the virulent genes of the newly isolated strain were simultaneously substituted by marker genes, which were subsequently excised using Cre/Lox system for vaccine safety concern. Notably, single cell FACS technology was applied to further promote virus purification efficiency. The combination of these state-of-art technologies greatly accelerated vaccine development. Finally, vaccination and challenge experiments proved this vaccine candidate’s protective efficacy in pigs and the promise to control current pseudorabies outbreak. This is, to our knowledge, the first successful vaccine development based on gene edit technologies, demonstrating these technologies leap from laboratory to industry. It may pave the way for future express antiviral vaccine development.