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関連する概念動画

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

795
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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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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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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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RNA Interference01:23

RNA Interference

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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
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Experimental RNAi02:15

Experimental RNAi

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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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関連する実験動画

Updated: Nov 6, 2025

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
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抗CRISPRタンパク質によるCRISPR-SpyCas9抑制の構造的基礎

De Dong1, Minghui Guo1, Sihan Wang1

  • 1HIT Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin 150080, China.

Nature
|April 28, 2017
PubMed
まとめ
この要約は機械生成です。

2つのアンチCRISPRタンパク質は,PAMを模倣し,DNA結合を阻害することで,Cas9を阻害する. この構造的な洞察は,Cas9の発展を可能にします.

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Substrate Generation for Endonucleases of CRISPR/Cas Systems
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科学分野:

  • 分子生物学
  • 構造生物学
  • 遺伝学

背景:

  • CRISPR-Cas9システムは 細菌の免疫メカニズムで 外来DNAを標的にします
  • Cas9酵素はガイドRNAを用いて特定の部位でDNAを割るため,プロトスペーサー付近モチーフ (PAM) が必要です.
  • 抗CRISPRタンパク質AcrIIA2とAcrIIA4はCas9を阻害するが,そのメカニズムは不明である.

研究 の 目的:

  • AcrIIA4がStreptococcus pyogenes Cas9 (SpyCas9) を抑制するメカニズムを解明する.
  • 制御可能なCas9システムを開発するための構造的基盤を提供すること.

主な方法:

  • シングルガイドRNA (sgRNA) とAcrIIA4を複合したSpyCas9のX線結晶学.
  • Cas9-sgRNA-AcrIIA4複合体の構造分析と比較

主要な成果:

  • AcrIIA4は sgRNAに依存した方法でSpyCas9と結合する.
  • AcrIIA4はPAMを模倣し,DNA結合部位を遮断することで,Cas9を阻害する.
  • また,AcrIIA4はCas9 RuvCの活性部位をシールドし,DNAの分裂を防ぐ.
  • sgRNA結合は,SpyCas9のAcrIIA4結合部位の形成を誘導する.

結論:

  • AcrIIA4は2つのメカニズムでSpyCas9を阻害する:DNA認識をブロックし,活性部位を遮断する.
  • この発見は,CRISPR-Cas9遺伝子編集ツールの"オフスイッチ"メカニズムを設計するための構造的基盤を提供します.
  • これらの相互作用を理解することは 精密なゲノム編集アプリケーションに不可欠です