Academy of Military Medical Sciences, Beijing 100850, China.
Institutes of Physical Science and Information Technology, Anhui University, Hefei 230000, China.
Int J Mol Sci. 2024 Oct 26;25(21):11514. doi: 10.3390/ijms252111514.
In recent years, there have been frequent global outbreaks of viral epidemics such as Zika, COVID-19, and monkeypox, which have had a huge impact on human health and society and have also spurred innovation in virus engineering technology. The rise of synthetic virus genome technology has provided researchers with a new platform to accelerate vaccine and drug development. Although DNA synthesis technology has made significant progress, the current virus genome synthesis technology still requires the assembly of short oligonucleotides of around 60 bp into kb-level lengths when constructing long segments, a process in which the commonly used polymerase chain reaction assembly (PCA) technology has high error rates and is cumbersome to operate. This study optimized the error correction conditions after PCA assembly, increasing the accuracy of synthesizing 1 kb DNA fragments from 4.2 ± 2.1% before error correction to 31.3 ± 3.1% after two rounds of correction, an improvement of over 6 times. This study provides a more efficient operational process for synthesizing virus genomes from scratch, indicating greater potential for virus engineering in epidemic prevention and control and the field of biomedicine.
近年来，全球频繁爆发寨卡、COVID-19 和猴痘等病毒疫情，对人类健康和社会造成了巨大影响，也推动了病毒工程技术的创新。合成病毒基因组技术的兴起为研究人员提供了一个新的平台，加速了疫苗和药物的开发。尽管 DNA 合成技术取得了重大进展，但目前的病毒基因组合成技术在构建长片段时仍需要将约 60bp 的短寡核苷酸组装成 kb 级长度，在这个过程中，常用的聚合酶链式反应组装（PCA）技术错误率高，操作繁琐。本研究优化了 PCA 组装后的纠错条件，将未经纠错时合成 1kb DNA 片段的准确率从 4.2±2.1%提高到两轮纠错后的 31.3±3.1%，提高了 6 倍以上。本研究为从头合成病毒基因组提供了更高效的操作流程，为病毒工程在疫情防控和生物医药领域的应用提供了更大的潜力。
