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相关概念视频

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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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

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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Conservative Site-specific Recombination and Phase Variation

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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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相关实验视频

Updated: Aug 26, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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克里斯普尔球状核酸

Chi Huang1, Zhenyu Han1, Michael Evangelopoulos2

  • 1Department of Chemistry and International Institute for Nanotechnology, Northwestern University, Evanston, Illinois 60208-3113, United States.

Journal of the American Chemical Society
|October 6, 2022
PubMed
概括
此摘要是机器生成的。

研究人员开发了新型的CRISPR球形核酸 (SNAs) 以有效编辑基因组. 这些Cas9 ProSNA增强了细胞传递和编辑效率, 无需严厉的方法, 扩大了CRISPR的应用.

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科学领域:

  • 生物技术
  • 分子生物学
  • 纳米技术

背景情况:

  • CRISPR/Cas9基因组编辑效率受阻于将组件输送到细胞和组织中的挑战.
  • 球状核酸 (SNA) 提供了增强细胞吸收的潜力,但尚未应用于基因编辑.

研究的目的:

  • 为改进基因组编辑设计和评估一类新的CRISPR SNA.
  • 增强CRISPR-Cas9组件的细胞传递和核定位.

主要方法:

  • 合成的Cas9 ProSNA:Cas9蛋白核与DNA外部密集修饰,预装有指导RNA.
  • 嵌入了GALA和核定位信号,以改善内体逃生和核向.
  • 在多个细胞系中评估了对蛋白酶消化的稳定性和基因组编辑效率.

主要成果:

  • 在没有电穿孔或转染剂的情况下,Cas9 ProSNAs表现出增强的细胞吸收.
  • 这些结构显示出对蛋白质酶降解的稳定性.
  • 在不同细胞系中实现基因组编辑效率32%至47%.

结论:

  • 新型CRISPR SNAs (Cas9 ProSNAs) 克服了基因组编辑的主要交付障碍.
  • 这些纳米结构显著提高了细胞吸收和编辑效率.
  • 这种进步有可能扩大CRISPR-Cas9技术的应用和影响.