Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragments.
Construction of an infectious horsepox virus vaccine from chemically synthesized DNA fragments.
复制标题
从化学合成的DNA片段中构建感染性的Horsepox病毒疫苗。
DOI：
10.1371/journal.pone.0188453
复制
发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Evans DH
中科院分区：
文献类型：
--
作者：
Noyce RS;Lederman S;Evans DH
Edward詹纳和他的同时代人认为他的天花疫苗起源于马，分子分析表明现代牛痘病毒（VACV）株与马痘病毒（HPXV）有共同的祖先。考虑到与现代VACV疫苗的毒性相关的问题，我们询问基于HPXV的疫苗是否可能提供上级替代方案。由于HPXV可能已经灭绝，并且已经鉴定的HPXV的唯一标本无法用于研究，因此我们探索了HPXV是否可以通过大规模基因合成获得。基于HPXV序列沿着两个157 nt VACV末端序列合成了10个大的（10-30 kb）DNA片段，并在用Shope纤维瘤病毒（SFV）感染的细胞中重组成活的合成嵌合HPXV（scHPXV）。对212 kbp scHPXV的测序证实其编码输入DNA的忠实拷贝。我们相信这是第一个使用合成生物学方法完整合成痘病毒的实验。这种scHPXV产生更小的噬斑，产生更少的细胞外病毒，并在小鼠中表现出比VACV更低的毒力，但仍然提供了针对致命VACV攻击的疫苗保护。总的来说，这些发现支持进一步开发scHPXV作为一种新的复制熟练的天花疫苗。
Edward Jenner and his contemporaries believed that his variolae vaccinae originated in horses and molecular analyses show that modern vaccinia virus (VACV) strains share common ancestry with horsepox virus (HPXV). Given concerns relating to the toxicity of modern VACV vaccines, we asked whether an HPXV-based vaccine might provide a superior alternative. Since HPXV may be extinct and the only specimen of HPXV that has been identified is unavailable for investigation, we explored whether HPXV could be obtained by large-scale gene synthesis. Ten large (10–30 kb) fragments of DNA were synthesized based on the HPXV sequence along with two 157 nt VACV terminal sequences, and were recombined into a live synthetic chimeric HPXV (scHPXV) in cells infected with Shope fibroma virus (SFV). Sequencing of the 212 kbp scHPXV confirmed it encoded a faithful copy of the input DNA. We believe this is the first complete synthesis of a poxvirus using synthetic biology approaches. This scHPXV produced smaller plaques, produced less extracellular virus and exhibited less virulence in mice than VACV, but still provided vaccine protection against a lethal VACV challenge. Collectively, these findings support further development of scHPXV as a novel replication-proficient smallpox vaccine.
登录
查看更多内容
影响因子：
5.4
作者：
Qin, Li;Evans, David H.
通讯作者：
Evans, David H.
影响因子：
5.4
作者：
Qin, Li;Upton, Chris;Evans, David H.
通讯作者：
Evans, David H.
影响因子：
5.4
作者：
DELANGE, AM;MCFADDEN, G
通讯作者：
MCFADDEN, G
影响因子：
3
作者：
Brodie, R;Smith, AJ;Upton, C
通讯作者：
Upton, C
影响因子：
5.4
作者：
Medaglia, Maria Luiza G.;Moussatche, Nissin;Damaso, Clarissa R.
通讯作者：
Damaso, Clarissa R.