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Updated: Jun 18, 2025
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
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在Mycolicibacterium neoaurum中使用Streptococcus thermophilus的Cas9进行高效的基因组工程
Gedan Xiang1, Tao Liu1, Lekai Li1
1State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.
Biotechnology letters
|July 31, 2024
概括
菌菌 (Mycolicibacterium neoaurum) 使用植物来生产类固醇. 一个新的CRISPR/sth1Cas9工具使M. neoaurum的快速基因工程成为可能,改善了工业类固醇中间体的合成.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 合成生物学 合成生物学
背景情况:
- 非致病性菌根细菌,如Mycolicibacterium neoaurum,自然地代谢植物来生产类固醇.
- M. neoaurum是工业合成类固醇药物中间体的关键生物,例如Pregnadien-20-carboxylic acid (PDC).
- 针对M. neoaurum的基因改造可以提高高价值类固醇产品的产量.
研究的目的:
- 为M. neoaurum. 开发一种简单而有效的基因工程策略.
- 创建一个可定制的CRISPR/sth1Cas9工具,用于M. neoaurum.精确的基因组编辑.
- 为了促进用于类固醇中间体合成的高产量工程菌株的生产.
主要方法:
- 构建一个可定制的等离子体工具 (pMSC9),集成sth1Cas9蛋白和sgRNA支架.
- 设计具有特定间隔序列的编辑等离子体,用于针对性基因破坏.
- 转化M. neoaurum,通过CRISPR/sth1Cas9诱导DNA双链断裂 (DSB),并通过NHEJ进行修复.
- 编辑等离子体的固化和使用无抗性培养方法的顺序基因删除.
主要成果:
- 成功开发和应用CRISPR/sth1Cas9系统用于M. neoaurum基因组编辑.
- 在M. neoaurum菌株中展示了快速和有针对性的基因删除.
- 创建适合进一步基因操纵和菌株改进的编辑M. neoaurum突变体.
结论:
- 克里斯普尔/sth1Cas9工具提供了一种快速有效的方法来编辑M. neoaurum.
- 这个系统显著增强了M. neoaurum. 的基因工程工具箱.
- 开发的工具支持通过改进的微生物工程来推进工业类固醇生产.
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