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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...
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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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Homologous Recombination02:31

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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

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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定向进化扩大了CRISPR-Cas12a的基因组编辑能力.

Enbo Ma1,2, Kai Chen1,2, Honglue Shi1,3

  • 1Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA 94720, United States.

Nucleic acids research
|July 17, 2025
PubMed
概括

研究人员设计了CRISPR-Cas12a (集群定期间隔的短Palindromic重复-Cas12a) 来识别新的DNA序列. 这种增强的基因组编辑工具Flex-Cas12a扩大了治疗和农业应用的准能力.

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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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科学领域:

  • 生物技术是生物技术.
  • 分子生物学分子生物学
  • 基因组学就是基因组学.

背景情况:

  • CRISPR-Cas12a是一种强大的RNA引导的基因组编辑系统.
  • 它的实用性受到一个狭窄的原始空间体相邻动机 (PAM) 识别 (5'-TTTV-3') 的限制,将向限制在基因组的1%左右.
  • 扩大PAM识别对于更广泛的基因组应用至关重要.

研究的目的:

  • 为了设计拉克诺斯皮拉氏菌Cas12a的变体,并扩展了PAM识别.
  • 为了克服野生类型Cas12a.的准限制.
  • 为了提高CRISPR-Cas12a的多功能性,用于基因组工程.

主要方法:

  • 基于细菌的定向进化试验.
  • 理性蛋白质工程的Cas12a.
  • 生物化学和基于细胞的测试来表征变异.
  • 对PAM识别特异性的分析.

主要成果:

  • 识别了具有扩展PAM识别的Cas12a变体,包括非正规的动机.
  • 开发了Flex-Cas12a,它除了正规的5'-TTTV-3' PAM之外还能识别5'-NYHV-3' PAM.
  • 扩展DNA识别部位到大约25%的人类基因组.
  • 证明保留了对正典PAM的认可.

结论:

  • 工程 Cas12a 变种,特别是 Flex-Cas12a,显著扩大基因组准能力.
  • Flex-Cas12a可以访问以前无法访问的基因组位置.
  • 这种进步为治疗和农业基因组工程提供了新的机会.