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相关概念视频

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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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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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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RNA Editing02:23

RNA Editing

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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相关实验视频

Updated: Feb 20, 2026

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

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使用CRISPR-Cas13进行RNA编辑

David B T Cox1,2,3,4,5,6, Jonathan S Gootenberg1,2,3,4,7, Omar O Abudayyeh1,2,3,4,6

  • 1Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.

Science (New York, N.Y.)
|October 27, 2017
PubMed
概括

研究人员使用CRISPR-Cas13和ADAR2开发了一个名为REPAIR的新型RNA编辑平台. 这种系统精确地纠正哺乳动物细胞的RNA水平的致病突变, 提供治疗潜力.

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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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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

Published on: May 25, 2018

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科学领域:

  • 生物技术
  • 分子生物学
  • 基因工程

背景情况:

  • 通过纠正与疾病相关的序列, 编辑RNA为治疗遗传疾病提供了一个有前途的策略.
  • 类型VI的CRISPR-Cas系统,特别是Cas13酶,是RNA引导的核酶,具有可编程RNA操纵的潜力.

研究的目的:

  • 设计一个Cas13的正确基因组,
  • 在哺乳动物细胞中使用催化无效的 Cas13 (dCas13) 来引导作用于 RNA 2 (ADAR2) 的腺脱氨酶进行RNA编辑.
  • 开发一个用于研究和治疗的多功能RNA编辑平台.

主要方法:

  • 对VI型CRISPR-Cas系统进行分析.
  • 设计一种催化无活性的Cas13 (dCas13) 变体.
  • 与作用于RNA 2 (ADAR2) 的腺脱氨酶一起表达dCas13,以进行向腺-至-氨酸 (A-to-I) RNA编辑.
  • 开发一种高特异性变体和最小化病毒传递系统.

主要成果:

  • 在使用REPAIR系统的哺乳动物细胞中成功编辑.
  • 在没有严格的序列约束的情况下, 展示了编辑具有致病突变的全长转录的能力.
  • 设计了一个高特异性变体和适合病毒传递的紧系统.

结论:

  • REPAIR系统是一个新的可编程RNA编辑平台.
  • 这项技术在基础研究,遗传疾病治疗开发和生物技术方面具有广泛的应用.
  • REPAIR提供了一种灵活的方法来纠正RNA序列,而无需对DNA进行修改.