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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

272
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...
272
CRISPR01:59

CRISPR

53.0K
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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Homologous Recombination02:31

Homologous Recombination

52.4K
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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相关实验视频

Updated: Sep 15, 2025

Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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一个基于CRISPR-Cas9的工具,用于剂量依赖的DNA损伤检测.

Valentyn Oksenych1, Pavlo Petakh2, Denis Kainov1

  • 1Department of Clinical and Molecular Medicine (IKOM), Norwegian University of Science and Technology, Trondheim, Norway.

The FEBS journal
|July 13, 2025
PubMed
概括

研究人员在酵母中创建了CRISPR-Cas9系统,以精确控制DNA双链断裂. 这种工具精确地研究了DNA损伤反应,并揭示了Tel1激酶局部化,为基因组稳定性研究提供了一个可扩展的平台.

关键词:
在这个过程中,我们得到了更多的信息.克里斯普尔-卡斯9是什么意思DNA 双链断裂发生这就是NHEJJ.电话电话电话电话电话同类的重组组合.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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科学领域:

  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.
  • 生物化学 生物化学

背景情况:

  • DNA双链断裂 (DSB) 是关键的DNA病变,可触发复杂的细胞反应.
  • 了解DNA损伤反应 (DDR) 对从癌症研究到衰老等领域至关重要.
  • 现有的诱导DSB的方法往往缺乏对中断频率和位置的精确控制.

研究的目的:

  • 开发一种基于CRISPR-Cas9的新型系统,用于精确和剂量依赖地诱导Saccharomyces cerevisiae中的DSB.
  • 在受控的DSB诱导后,研究关键DDR蛋白的动力学和局部化,例如Tel1激酶.
  • 建立一个可扩展的平台,用于研究不同生物体的基因组稳定性和DDR机制.

主要方法:

  • 利用CRISPR-Cas9系统准酵母中的Ty逆转移体,用于特定序列的DSB诱导.
  • 控制诱导的DSB数量 (×1,×15或×59) 以使剂量依赖性研究.
  • 采用显微镜技术观察 Tel1 激酶的局部化和焦点形成,以应对 DSBs.

主要成果:

  • 开发的系统允许精确,剂量依赖的DSB诱导,促进详细的DDR研究.
  • 观察到Tel1激酶局部化到核外围,并在DSB诱导时形成多个焦点.
  • 该研究确定了Cas9可用性在更高的破裂诱导水平上的局限性,为系统优化提供了洞察力.

结论:

  • 新的CRISPR-Cas9系统提供了一种强大而可扩展的工具,用于剖析DNA损伤反应途径.
  • 对DSB诱导的精确控制提高了研究DDR动态和蛋白质定位的能力.
  • 这个平台对于在各种模型生物中研究基因组稳定性和DDR机制具有广泛的适用性.