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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

CRISPR

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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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相关实验视频

Updated: Jul 25, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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经过工程改造的Staphylococcus auricularis Cas9具有高保真性

Nan Wei1,2, Lu Shang1,2, Jing Liu1,2

  • 1Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology
|June 30, 2023
PubMed
概括

设计的CRISPR-Cas9变体SauriCas9-HF1和SauriCas9-HF2显著提高了基因编辑的特异性. 这些改进的工具减少了非目标效应,扩大了CRISPR在研究和治疗中的应用.

关键词:
克里斯普尔-Cas9是什么意思耳性葡萄球菌Cas9 (SauriCas9) 是一种高保真度的高保真度显示器非目标效应的非目标效应

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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 生物技术是生物技术.

背景情况:

  • CRISPR-Cas9是一种强大的基因编辑工具,但非目标效应限制了其临床使用.
  • 耳性葡萄球菌Cas9 (SauriCas9) 是一个小的,活跃的Cas9 Ortholog,具有灵活的5'-NNGG-3' PAM识别.
  • 增强忠实性SaCas9 (efSaCas9) 通过单个突变显示出更好的特异性.

研究的目的:

  • 通过应用efSaCas9.9的突变来设计具有提高特异性的SauriCas9变体.
  • 评估工程SauriCas9变体的准特异性和非准效应.

主要方法:

  • 在SauriCas9和SaCas9.9之间的蛋白质序列对齐.
  • 具有特定突变的SauriCas9变体 (SauriCas9-HF1,SauriCas9-HF2) 的工程.
  • 评估使用目标深度测序和 GUIDE-seq. 的目标特异性的评估.

主要成果:

  • 两种设计的SauriCas9变种,即SauriCas9-HF1 (N269D) 和SauriCas9-HF2 (D270N),已经成功创建.
  • 与野生型SauriCas9.9相比,这两种变体都显示出明显改善的准特异性.
  • SauriCas9-HF2显示出非目标效应的显著减少 (某些部位的改善为61.6至111.9倍).

结论:

  • 设计的SauriCas9变体SauriCas9-HF1和SauriCas9-HF2增强了CRISPR-Cas9的特异性.
  • 这些变异减少了非目标突变,使得它们对精确的基因组编辑有价值.
  • 开发的SauriCas9变种扩展了CRISPR工具包,用于研究和治疗应用.