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相关概念视频

Genetic Screens02:46

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...

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Investigation of Genetic Dependencies Using CRISPR-Cas9-based Competition Assays
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使用幽灵细胞计量 (ghost cytometry) 进行高含量细胞表型的聚合CRISPR查.

Asako Tsubouchi1, Yuri An1, Yoko Kawamura1

  • 1ThinkCyte Inc., Tokyo 113-8654, Japan.

Cell reports methods
|March 26, 2024
PubMed
概括

这项研究引入了用于快速聚合CRISPR查的多式幽灵细胞计. 它可以基于光和无标签的表型进行高效的细胞分类,用于大规模的遗传分析.

关键词:
CP:生物技术 生物技术CP:成像成像的使用.细胞表型形成是细胞表型形成.流动细胞计量是流动细胞计量的方法.高含量细胞分析高含量细胞分析基于图像的细胞分类器没有标签的细胞分析.机器学习是机器学习.聚合了CRISPR查的情况.

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科学领域:

  • 细胞生物学 细胞生物学
  • 基因组学就是基因组学.
  • 生物技术是生物技术.

背景情况:

  • 基于图像的聚合CRISPR查推进了哺乳动物细胞中的基因型-表型映射.
  • 基于形态的快速细胞丰富是高含量查的一个挑战.
  • 现有的方法限制了各种细胞表型的大规模基因扰动查.

研究的目的:

  • 为了证明多式幽灵细胞计量用于快速聚合的CRISPR查.
  • 应用光和无标签的高含量表型用于细胞分类.
  • 克服查庞大的细胞群的局限性,以寻找特定的表型.

主要方法:

  • 采用多模式幽灵细胞计量与光和无标签模式.
  • 执行了针对酶的CRISPR选,使用光向RelA核转位.
  • 使用无标签模式对巨细胞两极化基因进行大规模查.

主要成果:

  • 成功执行了针对RelA转位的酶特异性CRISPR选.
  • 通过无标签查识别了参与巨细胞两极分化的基因.
  • 经过证明的无标签丰富,没有侵入性染色,保存细胞用于下游测定.

结论:

  • 多模式幽灵细胞计量使得跨多种表型的快速聚合CRISPR查成为可能.
  • 无标签的幽灵细胞计扩大了查潜力,特别是当标记物缺席时.
  • 这种方法保存了细胞,促进了进一步的下游生物测定.