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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 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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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: Jun 18, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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为CRISPR-Cas9基因编辑设计单循环MeV载体.

Ramya Rallabandi1, Brenna Sharp2, Spencer Majerus2

  • 1Mayo Clinic Graduate School of Biomedical Sciences, Virology and Gene Therapy Graduate Track, Mayo Clinic, Rochester, MN 55905, USA.

Molecular therapy. Methods & clinical development
|July 29, 2024
PubMed
概括

这项研究引入了一种麻疹病毒 (MeV) 载体,用于在人类细胞中高效的CRISPR-Cas9基因编辑. 这种新型载体使精确的基因淘汰和淘汰成为可能,为基因改造研究提供了一个灵活的平台.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?这就是Cas9的情况.这就是HDR HDR.这就是NHEJJ.基因编辑 基因编辑这就是 iPSC 的意义.麻疹是一种麻疹.病毒载体病毒载体

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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 病毒学 病毒学

背景情况:

  • 克里斯普尔-Cas9是一种强大的基因编辑工具,具有广泛的研究和治疗潜力.
  • 之前的工作确立了麻疹病毒 (MeV) 作为诱导多能干细胞生成的单周期重编程载体.

研究的目的:

  • 开发一种单循环MeV载体,用于将CRISPR-Cas9组件 (gRNA和Cas9核酶) 输入人体细胞.
  • 为了证明这个MeV向量的效率,用于精确的基因编辑,包括敲除和敲进.

主要方法:

  • 构建编码gRNA和Cas9核酶的单周期麻疹病毒 (MeV) 载体.
  • 将MeV载体传递给人类细胞,用于基因编辑应用.
  • 使用报告者和内源基因 (mCherry,HBB,FANCD1) 对目标编辑效率的评估.
  • 通过使用寡核酸捐赠体进行同质导向修复来评估精确的敲门.

主要成果:

  • 在人类细胞中,MeV 载体成功调解了目标基因编辑.
  • 对报告者 (mCherry) 和内源基因 (HBB,FANCD1) 均观察到有效的编辑.
  • 使用MeV载体和单链寡核酸供体实现了精确的基因敲定.

结论:

  • 开发的MeV载体作为一种新且可适应的基因淘汰和人类细胞中淘汰的平台.
  • 该系统有助于新兴基因编辑技术的整合.
  • 在基因编辑应用的基础和临床研究中,MeV向量显示出有前途的潜力.