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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/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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微DFBEST:一个dCas12b衍生的双功能基准编辑器,具有微生物遗传工程可编程编辑特性.

Wen-Liang Hao1, De-Zhi Geng2, Yu-Feng Liu3

  • 1Key Laboratory of Industrial Biotechnology (Ministry of Education), School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.

Synthetic and systems biotechnology
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概括

我们开发了MicroDFBEST,这是一款用于微生物的双功能基因编辑器,可以同时进行C-to-T和A-to-G基因组编辑. 这种多功能工具扩展了微生物基因编辑功能,用于合成生物学应用.

关键词:
这就是CRISPR/Cas12b.双功能基础编辑器遗传多样性 遗传多样性在现场进化的进化.有针对性的突变发生.

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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
  • 基因组编辑 基因组编辑

背景情况:

  • 现有的基础编辑器 (BEs) 在微生物应用方面存在局限性,包括受限制的突变发生和狭窄的编辑窗口.
  • 开发新的工具对于提高微生物的基因组编辑精度和多功能性至关重要.

研究的目的:

  • 为微生物设计了一种新的双功能基础编辑器 (DFBE),命名为MicroDFBEST.
  • 扩大微生物系统的基础编辑能力,用于各种合成生物学应用.

主要方法:

  • 融合高活性除氨酶 (evoCDA1和TadA9) 与一个核酶缺乏的Cas12b (dBhCas12b).
  • 使用 MicroDFBEST 在 26-33 nt 窗口内同时进行 C-to-T 和 A-to-G 编辑.
  • 通过修改融合蛋白表达和编辑代码来调整编辑特征.

主要成果:

  • 实现了微生物DFBEs报告的最广泛的编辑窗口.
  • 通过促进体多样化 (PylbP) 证明了灵活的基因表达调制.
  • 在本地基因组环境中通过突变热点扫描实现了向蛋白质进化.

结论:

  • MicroDFBEST提供了一个多功能平台,用于in situ基因调节,例如激活生物合成基因集群.
  • 该系统促进了蛋白质进化,包括底盘优化,具有广泛的合成生物学实用性.