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相关概念视频

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/Cas9 Genome Editing01:28

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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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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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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.
The recognition sites for Cre recombinase called LoxP...
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Prokaryotic Transcriptional Activators and Repressors01:58

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Prokaryotic Transcriptional Activators and Repressors01:58

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The organization of prokaryotic genes in their genome is notably different from that of eukaryotes. Prokaryotic genes are organized, such that the genes for proteins involved in the same biochemical process or function are located together in groups. This group of genes, along with their regulatory elements, are collectively known as an operon. The functional genes in an operon are transcribed together to give a single strand of mRNA known as polycistronic mRNA.
Transcription of prokaryotic...
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一个下一代平台,用于高度优化的CRISPR介导的转录抑制.

Andrew Kristof1, Krithika Karunakaran1, Yann Ferry2

  • 1School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA.

Journal of biotechnology
|September 26, 2025
PubMed
概括

研究人员设计了一种高级的CRISPR干扰 (CRISPRi) 系统,用于基因沉默. 这种新的dCas9-ZIM3-NID-MXD1-NLS抑制剂在各种应用中提供了增强的性能,改善了哺乳动物基因调节.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?在DCas9-抑制器聚变过程中.基因敲击下降的基因这就是MeCP2的意义.镇压器域名 镇压器域名转录的压制转录的压制

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科学领域:

  • 分子生物学分子生物学
  • 基因规则 基因规则
  • 蛋白质工程是指蛋白质工程.

背景情况:

  • 克里斯普尔干扰 (CRISPRi) 使用非活化的Cas9与抑制器域融合,用于基因沉默.
  • 目前的CRISPRi系统在不同的细胞系,基因点和指导RNA中显示出可变的疗效.

研究的目的:

  • 为增强哺乳动物基因调节开发一个高度优化的CRISPRi抑制剂.
  • 为了提高基于CRISPRi的基因沉默的一致性和有效性.

主要方法:

  • 通过切断抑制器域 (例如MeCP2) 和识别NID等最佳功能单元来进行蛋白质工程.
  • 组装和选用于新型压缩器的组合式多域融合库.
  • 优化核定位信号 (NLS) 配置,以提高敲击效率.

主要成果:

  • 一个超紧的NCoR/SMRT交互域 (NID) 截断增强了CRISPRi的淘汰效果,与正规域相比增加了40%左右.
  • 通过组合图书馆选,发现了四种新的抑制器融合.
  • 优化的NLS配置提高了压缩机敲击效率~50%.
  • 该dCas9-ZIM3-NID-MXD1-NLS系统证明了在多种细胞系和标中优越的基因沉默.

结论:

  • 新的dCas9-ZIM3-NID-MXD1-NLS系统代表了强大的基因沉默CRISPRi技术的重大进步.
  • 多域工程策略为开发下一代CRISPR工具提供了一个多功能框架.