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Mismatch Repair

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...

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相关实验视频

Updated: Jun 29, 2025

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

由基于dCas12a的双功能基编辑器驱动的多重复合的现场变异发生.

Yaokang Wu^1,2, Yang Li^1,2, Yanfeng Liu^1,2

¹Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.

Nucleic acids research

|April 3, 2024

概括

研究人员开发了MultiduBE,这是一种新的基础编辑器,可以在多个地点同时进行基因突变. 这种工具加速了对复杂生物功能的研究,并增强了对大肠杆菌和B. subtilis等细菌的代谢工程.

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

Published on: December 11, 2020

相关实验视频

Last Updated: Jun 29, 2025

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter

Published on: March 31, 2022

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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Published on: March 25, 2020

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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Published on: December 11, 2020

科学领域:

合成生物学合成生物学
分子生物学分子生物学
遗传学遗传学是一个

背景情况:

通过突变发生理解遗传多样性对于生物研究和工程至关重要.
目前用于组合在位突变发生的方法有限,这阻碍了对复杂生物功能的研究.

研究的目的:

设计和制造MultiduBE,一个基于dCas12a的多重复合双功能基数编辑器,用于组合在位突变发生.
为了证明MultiduBE在重编程细菌细胞生理学和代谢调节方面的有效性.

主要方法:

使用dCas12a开发MultiduBE,使用具有除氨酶活动的合成效应器 (duBE-1a,duBE-2b) 和合成分离器 (Sp4).
MultiduBE的应用在埃舍里奇亚大肠杆菌和细菌细菌中的多重现场突变发生.
鉴定一种链杆菌素耐药性突变和用于增强抗生素耐药性的组合性突变发生.

主要成果:

在大肠杆菌和B. subtilis.中成功实施多重化 in-situ 突变发生.
在B. subtilis.中发现一种新型的链杆菌素耐药性突变.
与单个突变对照组相比,组合突变显示出显著的改善:表面因子增加42%, рибофлавин标位增加15倍.

结论:

MultiduBE提供了一个方便而高效的平台,用于执行多重化的 in-situ 突变发生.
这项技术有助于复杂的遗传研究和代谢工程应用.
该工具能够快速生成具有理想特征的细菌菌株,例如增强抗生素耐药性和改善代谢物生产.