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CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

142
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...
142
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...
52.7K
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...
17.3K
Caspases01:24

Caspases

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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Updated: Aug 23, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

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対象外活動のための構造的基礎

Martin Pacesa1, Chun-Han Lin2, Antoine Cléry3

  • 1Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.

Cell
|October 28, 2022
PubMed
まとめ

CRISPR-Cas9によるゲノム編集は 意図しないDNA配列を標的とし 安全性の懸念を提起します 構造分析により,非正規の塩基配列とデレーションの収納により,標的外結合が可能になり,ガイドRNAの設計が改善された.

キーワード:
CRISPRについてカス9X線結晶学ベースペアリングゲノム編集ガイドRNA不一致核酵素ターゲット外

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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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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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Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
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Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

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関連する実験動画

Last Updated: Aug 23, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

805
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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Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
09:40

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

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科学分野:

  • 分子生物学
  • 遺伝学
  • 生物化学

背景:

  • CRISPRに関連した (Cas) 核酸 Cas9は強力なゲノム編集ツールです
  • Cas9の特異性は,誘導RNAと標的DNAの互換性に依存しています.
  • Cas9による標的外割れは,臨床応用において安全性のリスクを伴う.

研究 の 目的:

  • Cas9の標的外結合と分裂の構造的基礎を明らかにする.
  • 標的DNAの不一致と欠損を Cas9がどう処理するかを理解するために
  • より安全なCas9ベースのゲノム編集システムの合理的な設計に情報を提供するためです

主な方法:

  • Cas9-DNA複合体のX線結晶学
  • 異なる補完性を有する対象外基質の分析
  • オンターゲットとオフターゲットの結合モードの構造的比較

主要な成果:

  • Cas9は非正規の塩基配列の相互作用によって標的外DNAに結合する.
  • シングルヌクレオチドの削除は,塩基スキップまたは複数の非正規のペアによって対応されます.
  • PAM-ディスタル不一致は,Cas9のデュプレックス・アンペアリングとコンフォーマーション変化を誘導する.

結論:

  • 構造的な洞察が 標的外活動を説明する
  • 発見は,特異性を高めるためのガイドRNA設計の改善を容易にする.
  • この研究は,CRISPR技術のためのよりよいオフターゲットの予測アルゴリズムを開発するのに役立ちます.