一个DNA组装工具包,释放CRISPR/Cas9在代谢工程中的潜力
Tigran V Yuzbashev1,2, Evgeniya Y Yuzbasheva3, Olga E Melkina4
1Department of Bioengineering, Imperial College London, London, SW7 2AZ, UK. t.yuzbashev@gmail.com.
Communications biology
|August 18, 2023
概括
改进了CRISPR/Cas9基因编辑工具包,使微生物细胞工程更容易. 新方法简化了DNA集成和指导RNA组装,增强了代谢工程应用.
科学领域:
- 微生物生物技术 微生物生物技术
- 合成生物学 合成生物学
- 基因编辑技术 基因编辑技术
背景情况:
- 在微生物菌株工程中,CRISPR/Cas9使得无标记DNA集成成为可能,简化了以前的方法.
- 目前的CRISPR/Cas9工具包在组装和实施方面存在挑战,限制了更广泛的可访问性.
- 高效的微生物细胞工程对于生物技术和合成生物学的进步至关重要.
研究的目的:
- 为了解决目前用于微生物工程的CRISPR/Cas9工具包的局限性.
- 开发改进的,用户友好的CRISPR/Cas9系统,以提高可访问性和实施性.
- 为高效的代谢工程应用创建一个全面的工具包.
主要方法:
- 开发了一个用于在无标记和基于标记的DNA整合结构之间切换的系统.
- 启用了使用金门组件将多基因整合录音带重定向到替代基因组位点.
- 通过重组建立了一个快速的in-vivo方法,通过重组组合组装指导RNA序列.
- 将这些方法整合到一个全面的工具包中,以YaliCraft工具包为Yarrowia lipolytica的例子.
主要成果:
- 为Yarrowia lipolytica创建了一个多功能工具包 (YaliCraft),包括147个质粒和7个模块.
- 成功生成并描述了137个推动者的图书馆.
- 构建了一种能够合成373.8mg/L同质酸的新菌株.
结论:
- 开发的CRISPR/Cas9工具包显著提高了微生物细胞工程的易用性和效率.
- 这些改进有助于在代谢工程和合成生物学中更广泛采用CRISPR/Cas9技术.
- 这项工作为构建各种应用的工程微生物菌株提供了一个强大的平台.
相关概念视频
01:28CRISPR/Cas9 Genome Editing
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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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02:53CRISPR and crRNAs
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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