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相关概念视频

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.6K
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...
1.6K
CRISPR01:59

CRISPR

57.4K
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...
57.4K
Homologous Recombination02:31

Homologous Recombination

62.5K
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...
62.5K
CRISPR and crRNAs02:53

CRISPR and crRNAs

18.7K
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...
18.7K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.6K
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...
6.6K

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相关实验视频

Updated: Jan 12, 2026

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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计算方法设计Cas蛋白质,以实现高效的基因组编辑.

Muhammad Qaiser Fatmi1, Aneeqa Nadeem2, Mehraj Abbasov3

  • 1Department of Biosciences, COMSATS University Islamabad, Islamabad, Pakistan. qaiser.fatmi@comsats.edu.pk.

Methods in molecular biology (Clifton, N.J.)
|November 1, 2025
PubMed
概括
此摘要是机器生成的。

研究人员现在可以理性地设计改进的CRISPR/Cas基因组编辑工具. 该协议通过使用计算方法提高了Cas蛋白的稳定性和特异性,以实现更精确的DNA向.

关键词:
这就是CRISPR/Cas9的作用.共同进化的合.基因组编辑 基因组编辑分子动力学模拟模型突变的稳定性预测预测网络的中心性 网络的中心性蛋白质工程是一种蛋白质工程.

更多相关视频

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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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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相关实验视频

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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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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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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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科学领域:

  • 生物化学 生物化学
  • 分子生物学分子生物学
  • 生物信息学是一种生物信息学.

背景情况:

  • 克里斯普尔/卡斯系统使可编程的,特定序列的DNA定位成为基因组编辑的目标.
  • 由于非目标效应和可变效率,广泛应用受到阻碍.
  • 需要合理的设计策略来设计增强的CRISPR/Cas变体.

研究的目的:

  • 为设计改进的CRISPR/Cas变体提供一个集成的计算协议.
  • 为了提高Cas蛋白的稳定性和特异性,用于基因组编辑.
  • 为研究人员提供简单易行的合理蛋白质工程步骤.

主要方法:

  • 同进化合分析以确定关键残留物.
  • 对能量有利的替代物进行突变稳定性预测.
  • 网络中心性的分析,以评估对分子内通信的影响.
  • 网络分析和分子动力学 (MD) 模拟的代应用.
  • 使用MD模拟验证结构完整性和动态行为.

主要成果:

  • 鉴定对Cas蛋白功能至关重要的保存和共变残留物.
  • 预测增强蛋白质稳定性的突变,同时保持全相互作用.
  • 评估突变对分子内通讯路径的影响.
  • 通过MD模拟验证工程变体的结构完整性和动态行为.

结论:

  • 集成的多尺度计算策略简化了优化 Cas 蛋白质的工程.
  • 这种方法促进了CRISPR/Cas变体的合理设计,提高了稳定性和特异性.
  • 该协议使研究人员能够开发更精确,更有效的基因组编辑工具.