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相关概念视频

Genetic Screens02:46

Genetic Screens

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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...
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Genome-scale functional mapping of the mammalian whole brain with in vivo Perturb-seq.

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Competitive Genomic Screens of Barcoded Yeast Libraries
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在体内大规模并行进行 Perturb-seq 查.

Xinhe Zheng1, Patrick C Thompson1, Cassandra M White1

  • 1Department of Neuroscience, Dorris Neuroscience Center, Scripps Research, La Jolla, CA, USA.

Nature protocols
|February 12, 2025
PubMed
概括

这项研究介绍了活体Perturb-seq,一种使用基因编辑和单细胞基因组学的方法,以发现完整组织中的基因功能. 该协议指导研究人员在疾病研究中剖析基因作用和细胞特异性作用.

科学领域:

  • 基因组学就是基因组学.
  • 分子生物学分子生物学
  • 系统生物学 系统生物学

背景情况:

  • 基因组研究已经确定了许多疾病风险基因,但它们的功能和机制在很大程度上是未知的.
  • 了解完整组织内特定细胞类型的基因功能对于推进疾病研究至关重要.
  • 在体内剖析基因功能和细胞类型特异性的方法中存在差距.

研究的目的:

  • 描述一个详细的协议,以执行大量并行 in vivo Perturb-seq.
  • 为功能基因组学设计和执行体内Perturb-seq实验提供指导.
  • 为了促进采用体内Perturb-seq用于各种生物学问题.

主要方法:

  • 利用最先进的基因编辑工具进行可编程的基因扰动.
  • 采用高内容,高分辨率的单细胞基因组测试用于表型读取.
  • 开发了一种使用多功能腺相关病毒 (AAV) 载体进行体内输送的协议.

主要成果:

  • 详细的逐步程序,从扰乱库设计到AAV生产和管理.
  • 突出了对实验成功的关键质量控制检查点.
  • 讨论了 in vivo Perturb-seq. 的数据分析选项和可用的软件.

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结论:

  • 在体内Perturb-seq提供了一种强大的方法来加速哺乳动物系统中的功能基因组学研究.
  • 该协议使研究人员能够在体内研究基因功能和细胞特异性作用.
  • 这种方法有可能回答广泛的生物学问题,进步我们对健康和疾病的理解.