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相关概念视频

CRISPR01:59

CRISPR

50.9K
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...
50.9K
CRISPR and crRNAs02:53

CRISPR and crRNAs

17.0K
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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相关实验视频

Updated: Jun 30, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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在多态体Ciona intestinalis中优化CRISPR/Cas9方法

Alessandro Pennati1, Miloš Jakobi1, Fan Zeng1

  • 1Institute of Zoology, University of Innsbruck, 6020, Innsbruck, Austria; Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, 6020, Innsbruck, Austria.

Developmental biology
|March 15, 2024
PubMed
概括

我们通过改进指导RNA设计,验证体内测试,并增强体内传递来优化Ciona intestinalis模型生物体中的CRISPR/Cas9基因编辑. 这些进步提高了CRISPR淘汰效率,并可转移到其他物种.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?第9个案例:双胞胎这就是Ciona intestinalis.多态性多态性多态性突尼西亚国家 (Tunicates)

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Last Updated: Jun 30, 2025

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Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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科学领域:

  • * 发育生物学 发育生物学
  • * 分子生物学 * 分子生物学
  • * 遗传学 在遗传学方面

背景情况:

  • * Ciona intestinalis是脊椎动物发育研究的关键无脊椎动物模型,因为它具有共同的特征和实验优势.
  • * 基因多态度高和Ciona的马赛克表达阻碍了有效的CRISPR/Cas9淘汰.
  • * 为了减少动物使用和克服Ciona研究中的季节性挑战,需要采用体外方法.

研究的目的:

  • * 优化Ciona intestinalis中的CRISPR/Cas9基因编辑协议,以提高效率和更广泛的适用性.
  • * 开发出in silico,in vitro和in vivo策略,以获得更可靠的CRISPR淘汰.
  • * 增强Ciona作为遗传研究模型生物的实用性.

主要方法:

  • * In silico:通过对目标区域进行测序并与参考基因组对齐来纠正指导RNA (sgRNA) 设计,以考虑亚种多态性.
  • *体外:使用体外测试验证sgRNA效率,以减少体内实验.
  • *在体内:将Cas9与Cas9:Geminin进行比较,以提高编辑效率,利用下一代测序 (NGS) 进行基因型定型,并使用双重CRISPR进行大规模删除.

主要成果:

  • *优化的sgRNA设计和in silico分析通过纠正Ciona亚种多态性来提高准确性.
  • *在体外测试显示适合 sgRNA 验证,但也突出了不匹配耐受性.
  • *Cas9:Geminin提高了体内编辑效率,使得可以访问早期表达的基因.
  • *下一代测序简化了有效的sgRNAs的识别,双CRISPR产生了显著的删除.

结论:

  • * 提出的改进显著提高CRISPR/Cas9在Ciona intestinalis中的效率和可靠性.
  • *这些优化的方法有助于在新的研究环境中建立CRISPR技术,并可转移到其他生物体.
  • *这项研究为推进基因工程在Ciona及其他地区的发展提供了坚实的框架.