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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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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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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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相关实验视频

Updated: Jul 25, 2025

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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利用定向进化来询问和优化CRISPR/Cas指导RNA架构.

Korie Bush1, Giulia I Corsi2, Amy C Yan3

  • 1Department of Surgery, Duke University, Durham, NC 27710, USA; University Program in Genetics and Genomics, Duke University, Durham, NC 27710, USA; Moderna Genomics, Cambridge, MA 02139, USA.

Cell chemical biology
|June 30, 2023
PubMed
概括
此摘要是机器生成的。

研究人员开发了一种新方法,BLADE,以创建改进的CRISPR-Cas9基因编辑工具. 这种方法通过进化新的单向导RNA变异来增强编辑困难的DNA序列.

关键词:
克里斯普尔是什么意思?克里斯普尔是什么意思?这就是Cas9的情况.编辑DNA的编辑这就是SELEXEX.一个aptamer的应用程序.导向RNA指导RNA是指导RNA的指导RNA.分子进化分子演变.

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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相关实验视频

Last Updated: Jul 25, 2025

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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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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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:

  • 基于CRISPR的基因编辑已使基因组工程取得了先进的进展.
  • 由于单向导RNA (sgRNA) 和Cas9复合体内的不富有成效的相互作用,某些DNA序列仍然难以准.

研究的目的:

  • 开发一种用于识别多种sgRNA变异的新方法,以提高CRISPR-Cas9编辑效率.
  • 为了克服针对基因组工程具有挑战性的DNA序列的局限性.

主要方法:

  • 采用一种功能性的SELEX (通过指数式丰富的连接体的系统进化) 方法,称为BLADE (结合和连接体激活的定向进化).
  • BLADE被用来识别和进化有效结合Streptococcus pyogenes Cas9并支持DNA裂变的sgRNA变体.

主要成果:

  • 确定了许多具有显著序列可形性的sgRNA变异.
  • 特定的sgRNA变体与特定的DNA结合反意义域展现了增强的合作伙伴关系,从而提高了各种目标站点的编辑效率.

结论:

  • 使用BLADE方法的分子进化可以产生能够高效编辑具有挑战性的DNA序列的CRISPR-Cas9系统.
  • 这种选择策略为创建具有广泛活动的sgRNA提供了有价值的工具,使基因组更容易接受工程.