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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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CRISPR01:59

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CRISPR/Cas9 Genome Editing01:28

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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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CRISPR and crRNAs02:53

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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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通过CRISPR/Cas9进行可编程RNA识别和分离.

Mitchell R O'Connell1, Benjamin L Oakes1, Samuel H Sternberg2

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.

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概括

现在,CRISPR-Cas9可以针对RNA,而不仅仅是DNA. 通过使用单独的DNA序列 (PAMmer),Cas9可以结合和切割特定的RNA分子,从而实现可编程的转录识别.

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科学领域:

  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.
  • 生物技术是生物技术.

背景情况:

  • 克里斯普尔相关蛋白9 (Cas9) 是一种DNA内核酶,通过RNA互补性和原空间体相邻动机 (PAM) 引导目标识别.
  • Cas9被广泛用于基因组工程,但被认为无法向RNA.
  • 了解Cas9的基质特异性对于扩大其应用至关重要.

研究的目的:

  • 为了调查Cas9是否可以通过PAM序列的帮助来准单链RNA (ssRNA).
  • 探索Cas9-RNA相互作用和裂变的机制.
  • 为了证明Cas9在生物学上对RNA向的实用性.

主要方法:

  • 在体外测试中使用Cas9,指导RNA,ssRNA点和转呈的PAM寡核酸 (PAMmers).
  • 用PAMmers对ssRNA标的Cas9结合亲和力的表征.
  • 通过PAMmers刺激的特定位点ssRNA裂变的证明.
  • 从细胞中分离内源信使RNA (mRNA) 的方法的应用.

主要成果:

  • 当PAM序列通过DNA寡核化物 (PAMmer) 提供时,Cas9与ssRNA标具有高度亲和力.
  • PAM 激素刺激了Cas9介导的,网站特定的ssRNA点的内核解裂.
  • 使用专门设计的PAMmers,Cas9可以被导向RNA目标,同时避免使用DNA序列.
  • 这一策略成功地使得从细胞中分离出特定的内源mRNA.

结论:

  • Cas9的PAM结合基本上与其基质选择有关,使RNA向成为可能.
  • Cas9可以被编程为特定的转录识别和分离,而不需要目标标签.
  • 这项工作将Cas9的实用性扩展到DNA之外,为RNA操纵和分析开辟了新的途径.