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相关概念视频

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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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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相关实验视频

Updated: May 28, 2025

Ubiquitous and Tissue-specific RNA Targeting in Drosophila Melanogaster using CRISPR/CasRx
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Ubiquitous and Tissue-specific RNA Targeting in Drosophila Melanogaster using CRISPR/CasRx

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使用CRISPR-Cas13d精确准RNA的方法

Sydney K Hart1,2, Simon Müller1,2, Hans-Hermann Wessels1,2

  • 1New York Genome Center, New York, NY, USA.

Nature biotechnology
|February 11, 2025
PubMed
概括
此摘要是机器生成的。

通过使用低RfxCas13d表达来缓解CRISPR-Cas13RNA降解问题,从而实现精确的转录组编辑. 一种高忠实度的Cas13变体显示了降低的附带活性,可能是由于较低的核酶功能.

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CRISPR Gene Editing Tool for MicroRNA Cluster Network Analysis

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相关实验视频

Last Updated: May 28, 2025

Ubiquitous and Tissue-specific RNA Targeting in Drosophila Melanogaster using CRISPR/CasRx
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科学领域:

  • 分子生物学分子生物学
  • 基因编辑技术的技术
  • 在RNA生物学,RNA生物学.

背景情况:

  • 克里斯普尔-Cas13系统提供精确的RNA向,但引起了人们对不必要的附带RNA降解的担忧.
  • 这种附带活动可能会扰乱转录组并限制治疗应用.

研究的目的:

  • 研究RfxCas13d表达水平与附带RNA降解之间的关系.
  • 开发使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.使用CRISPR-Cas13.
  • 探索高保真Cas13变体中减少抵押活动背后的机制.

主要方法:

  • 采用了转录组规模和组合组合选方法.
  • 系统地改变了RfxCas13d表达水平.
  • 分析了高保真性Cas13变体的核酶活性和副作用.

主要成果:

  • 附带RNA降解仅在高RfxCas13d表达水平时观察到.
  • 低复制RfxCas13d在屏幕上没有显著的附带活动的情况下实现了高目标敲击.
  • 高保真性Cas13变异体现出降低的附带活性,与整体核酶能力的降低相关.

结论:

  • 在CRISPR-Cas13应用中,优化RfxCas13d表达对于最小化附带RNA降解至关重要.
  • 低表达RfxCas13d和高保真性变体对安全有效的转录组全方位RNA编辑和治疗用途充满希望.
  • 减少核酶能力似乎是缓解Cas13系统附带活动的关键因素.