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関連する概念動画

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

2.6K
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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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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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

65.4K
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

19.5K
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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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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精密性が向上した合理的に設計されたCas9核酸

Ian M Slaymaker1, Linyi Gao2, Bernd Zetsche1

  • 1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA. Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Science (New York, N.Y.)
|December 3, 2015
PubMed
まとめ
この要約は機械生成です。

強化特異性Cas9 (eSpCas9) と呼ばれる Cas9の改造された変種は,ゲノム編集における意図しないDNA分裂を大幅に減少させます. これらの改良されたCas9ツールは より安全な遺伝子編集アプリケーションのために 精密なターゲティングを維持します

さらに関連する動画

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

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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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関連する実験動画

Last Updated: Mar 29, 2026

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
11:37

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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科学分野:

  • 分子生物学
  • バイオテクノロジー
  • ゲノミクス

背景:

  • RNA誘導内核酵素Cas9は,標的型DNA改変を可能にするゲノム編集の重要なツールです.
  • Cas9酵素は,RNA分子によって導かれる特定のゲノム部位で二重鎖断裂 (DSB) を生み出します.
  • Cas9の重要な制限は,編集の正確性と安全性を損なう,標的外部を割る可能性である.

研究 の 目的:

  • 強化された特異性を持つCas9 (eSpCas9) 変種をStreptococcus pyogenes Cas9 (SpCas9) から生成する.
  • 人体細胞におけるeSpCas9変種の特異性と標的割れ活性を厳密に評価する.
  • ゲノムエディティングアプリケーションの 対象外割れの問題に対処するためです

主な方法:

  • SpCas9を修正するために,構造誘導タンパク質工学が採用されました.
  • 対象外部位におけるCas9媒介のDNA分裂を検出するために,標的型深層配列解析を用いた.
  • Cas9の全ゲノム活性を包括的に評価するために,偏りのない全ゲノムオフターゲットの分析が行われました.

主要な成果:

  • 設計されたeSpCas9の変種は,ワイルド型SpCas9と比較して,標的外DNAの分裂を大幅に減少させました.
  • eSpCas9のバリエーションは,高いターゲット割れ効率を維持し,必要な場所での正確な編集を保証しました.
  • 標的型と全ゲノム分析の両方が,eSpCas9の多様性の強化を確認した.

結論:

  • 強化された特異性SpCas9 (eSpCas9) 変種は,ゲノム編集におけるオフターゲット効果を効果的に最小限に抑えます.
  • eSpCas9は強力な標的割れ活動を保持しており,正確な遺伝子編集のための信頼できるツールです.
  • これらのCas9変種は,高特異性を要求する多様なゲノム編集アプリケーションにおいて,安全性と有効性を向上させています.