用CRISPR系统对Acidithiobacillus ferridurans进行基因工程,以减轻生物开采中的有毒释放
Jinjin Chen1,2, Yilan Liu1,2, Radhakrishnan Mahadevan1,3
1Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario M5S 3E5, Canada.
Environmental science & technology
|August 9, 2023
概括
合成生物学增强生物采矿使用CRISPR-Cas9基因编辑在Acidithiobacillus细菌. 这提高了金属提取效率,同时减少了酸和铁生产对环境的影响.
科学领域:
- 微生物生物技术和合成生物学
- 环境生物技术和生物修复
- 生物洗和生物采矿应用.
背景情况:
- 生物采矿使用像Acidithiobacillus这样的微生物来提取金属,但过度的酸和铁生产会导致环境问题.
- 缓慢生长的Acidithiobacillus菌株的基因工程很困难,阻碍了可持续的生物采矿进步.
- 合成生物学为改善Acidithiobacillus菌株提供了潜在的解决方案,以实现环保和成本效益的生物采矿.
研究的目的:
- 为生物开采细菌Acidithiobacillus ferridurans JAGS建立和优化基于CRISPR的基因编辑工具.
- 设计A. ferridurans JAGS以提高生物采矿性能,减少环境副产品.
- 为了证明工程菌株在从铁矿尾矿中提取的实用性.
主要方法:
- 开发一种CRISPR-dCas9系统,用于A. ferridurans JAGS的基因淘汰,针对硫氧化基因.
- 构建一个全集CRISPR-Cas9系统,用于高效的基因组编辑,包括基因删除,促进物替代和基因插入.
- 创建一个双编辑的HdrB-Rus菌株和随后的生物开采实验,使用皮洛矿尾矿.
主要成果:
- 通过使用CRISPR-dCas9.9成功下调硫氧化基因.
- 通过CRISPR-Cas9系统实现了高效的基因删除 (HdrB3),促进子替代 (Prus到Ptac) 和GFP插入.
- 工程菌株显示与野生类型相似的Ni回收,但在液中铁和硫酸显著减少.
结论:
- 在A. ferridurans JAGS.中建立了有效的CRISPR-dCas9和CRISPR-Cas9系统用于基因操纵.
- 证明了合成生物学在开发强大的和环保的生物矿业微生物中的潜力.
- 开发的工具对于微生物基因功能和合成生物学辅助生物采矿的未来研究非常有价值.
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