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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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Conservative Site-specific Recombination and Phase Variation02:53

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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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适应CRISPR相关的转基因用于快速和高通量逆遗传学.

David W Basta1,2,3, Franz G Zingl2,3, Yiyan Yang4

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概括

通过使用CRISPR关联转体子 (CAST) 和指导RNA条形码,MultiCAST可实现高通量细菌遗传查. 这个平台简化了基因组工程,并有效地识别了特定条件的健身基因.

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科学领域:

  • 分子生物学分子生物学
  • 基因组学就是基因组学.
  • 合成生物学 合成生物学

背景情况:

  • 与CRISPR相关的转子子 (CAST) 是细菌基因组工程的可编程工具.
  • 现有的CAST系统没有针对高通量遗传查应用进行优化.

研究的目的:

  • 开发MultiCAST,一个简化的平台,用于快速,可扩展,指导RNA导向的转子子插入细菌.
  • 为了实现聚合,高通量基因选,使用片序列测序进行分子条形编码.
  • 确定影响CAST转化效率的因素,并优化选协议.

主要方法:

  • 结合性输送的等离子体编码CAST机械和一个迷你转子与指导RNA.
  • 使用指导RNA序列作为分子条形码进行聚合查.
  • 开发一种机器学习模型,根据目标序列特征来预测活性导向RNA.
  • 研究核相关蛋白H-NS在CAST活动中的作用.

主要成果:

  • 多重CAST可实现单步,针对性转子子插入,用于可扩展的遗传选.
  • 优化的结合比可以最大限度地减少导向-转位子交叉声.
  • 机器学习模型准确地预测了活性导向RNA.
  • H-NS被确定为CAST活动的抑制剂,解释了可变插入频率.
  • 在大肠杆菌中进行的大规模查,确定了特定条件的适应性基因.

结论:

  • MultiCAST显著提高了细菌基因组规模功能屏幕的可访问性,速度和吞吐量.
  • 该平台有助于识别具有特定条件健身效应的基因.
  • MultiCAST可以适应各种细菌物种,扩大其在研究和生物技术中的应用.