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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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CRISPR01:59

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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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Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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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.
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相关实验视频

Updated: Sep 12, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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没有序列限制的CRISPR/Cas系统引导的等离子体突变发生.

Fengjiao Zhao1, Feng Chen1, Huahang Yu1

  • 1Institute of Analytical Chemistry and Instrument for Life Science, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.

Fundamental research
|August 8, 2025
PubMed
概括
此摘要是机器生成的。

这项研究介绍了CRISPR/Cas系统引导的等离子体突变发生,用于工程蛋白质变体. 新方法有效地在等离子体上创建用户定义的突变库,改进蛋白质工程和基因组编辑工具.

关键词:
这是CRISPR/Cas系统.DNA组装组件的组装等离子体突变发生在等离子体中.蛋白质工程是一种蛋白质工程.序列可编程性 序列可编程性

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科学领域:

  • 分子生物学分子生物学
  • 生物技术是生物技术.
  • 蛋白质工程是指蛋白质工程.

背景情况:

  • 等离子体突变发生对于设计具有新功能的蛋白质至关重要.
  • 目前的方法,如基于PCR的技术和亚克隆具有局限性,特别是在产生某些突变发生的策略所需的单链圆形等离子体.
  • 在等离子体上直接产生突变提供了优势,但面临着序列限制.

研究的目的:

  • 开发一种新的CRISPR/Cas系统导向方法用于等离子体突变发生.
  • 为了实现在等离子体上直接生成用户定义的突变库,克服现有的序列限制.
  • 为了改进基因组编辑蛋白FnCpf1变体,改进其特性.

主要方法:

  • 使用CRISPR/Cas系统与导向RNA (gRNA) 和Cas9尼克酶生成单链圆形质粒.
  • 使用这些单链等离子体作为直接突变发生的模板.
  • 将突变发生法与蛋白质工程的理性设计原则相结合.

主要成果:

  • 通过CRISPR/Cas系统导向突变发生法,在等离子体上成功生成了用户定义的突变库.
  • 该方法证明了广泛的序列可编程性和与甲基化塑料的兼容性.
  • 工程FnCpf1变种显示扩大了PAM范围,减少了目标外效应,增强了基因组编辑特异性.

结论:

  • CRISPR/Cas系统引导的等离子体突变发生是一种有效的工具,用于创建突变库和工程蛋白质.
  • 这种方法克服了与以前的等离子体突变发生技术相关的序列限制.
  • 工程FnCpf1变种提供了改进的基因组编辑功能,松开了PAM约束,增加了目标特异性.