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相关概念视频

CRISPR01:59

CRISPR

57.1K
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...
57.1K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.4K
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...
1.4K
CRISPR and crRNAs02:53

CRISPR and crRNAs

18.5K
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...
18.5K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.5K
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...
6.5K
Homologous Recombination02:31

Homologous Recombination

61.9K
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...
61.9K

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相关实验视频

Updated: Dec 18, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

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非常快的CRISPR按需使用

Yang Liu1, Roger S Zou2, Shuaixin He3

  • 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University, Baltimore, MD, USA. tjha@jhu.edu bwu20@jhu.edu.

Science (New York, N.Y.)
|June 13, 2020
PubMed
概括
此摘要是机器生成的。

我们开发了一种非常快速的CRISPR (vfCRISPR) 光激活基因组编辑工具. 这种方法精确地控制DNA双链断裂,用于DNA修复动态的高分辨率研究.

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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相关实验视频

Last Updated: Dec 18, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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科学领域:

  • 分子生物学
  • 遗传学
  • 生物技术

背景情况:

  • CRISPR-Cas系统是一种强大的基因组编辑工具.
  • 对于DNA修复机制的详细研究,需要对DNA裂变进行精确的时间控制.

研究的目的:

  • 开发一种可导光的CRISPR-Cas9系统,用于DNA修复的高分辨率时空分析.
  • 研究DNA双链断裂的动力学和分子事件.

主要方法:

  • 开发了一种用于光激活Cas9结合和分裂的封闭RNA策略 (vfCRISPR).
  • 使用单细胞光成像来监测DNA修复焦点 (例如53BP1) 和蛋白质动态 (例如MRE11).
  • 测量了H2AX酸化的传播作为DNA损伤反应的标志物.

主要成果:

  • vfCRISPR 能够在二次和亚微米尺度上快速,同步地诱导 DNA 双链断裂.
  • 在几分钟内观察到细胞对DNA损伤的反应,包括MRE11保留后的结合.
  • 描述了H2AX化扩散的动力学和53BP1修复焦点的动力学.
  • 通过成像指导进行单个基因组操纵.

结论:

  • 在研究DNA修复方面提供了前所未有的时空分辨率.
  • 该系统允许对DNA损伤反应途径进行详细的动力分析.
  • 允许精确的基因组操纵进行先进的生物研究.