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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/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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Homologous Recombination02:31

Homologous Recombination

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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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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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Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

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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

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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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使用CRISPR-Cas13进行RNA向

Omar O Abudayyeh1,2,3,4,5, Jonathan S Gootenberg1,2,3,4,6, Patrick Essletzbichler1,2,3,4

  • 1Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA.

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概括

科学家设计了CRISPR-Cas13a以精确操纵哺乳动物细胞中的RNA. 这种新工具提供了针对性的RNA淘汰和活细胞转录跟踪,比现有方法具有更高的特异性.

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科学领域:

  • 分子生物学
  • 遗传学
  • 生物技术

背景情况:

  • 用于RNA操纵和测量的分子工具是有限的.
  • 像RNA干扰这样的现有方法会产生异常效应.
  • 可视化RNA通常需要外源标签.

研究的目的:

  • 设计CRISPR-Cas13a系统用于哺乳动物细胞中的RNA敲除和结合.
  • 评估LwaCas13a针对RNA的疗效和特异性.
  • 开发一个可编程的平台来研究活细胞中的RNA.

主要方法:

  • 选了15名Cas13a骨科医生,以确定最有效的变种 (LwaCas13a).
  • 在哺乳动物和植物细胞中异质表达LwaCas13a.
  • 用于催化无活性的LwaCas13a进行RNA结合和跟踪.

主要成果:

  • LwaCas13a 证明了对报告者和内源性转录的有效制.
  • 达到与RNA干扰相比较的降解水平,但具有更高的特异性.
  • 使用非活性LwaCas13a在活细胞中的可编程追踪.

结论:

  • CRISPR-Cas13a是一个用于哺乳动物细胞RNA研究的多功能平台.
  • LwaCas13a提供了一种特定且可编程的RNA敲击和可视化工具.
  • 这项技术在治疗RNA相关疾病方面具有潜在的应用.