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相关概念视频

Somatic to iPS Cell Reprogramming01:29

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Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012...
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Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for...
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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
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相关实验视频

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通过近距离依赖生物化研究细胞动力学:体细胞重编程.

Reuben Samson1,2, Francesco Zangari1,2, Anne-Claude Gingras3,4

  • 1Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, ON, Canada.

Methods in molecular biology (Clifton, N.J.)
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概括
此摘要是机器生成的。

本研究介绍了时间解析的靠近依赖生物化 (PDB) 协议,用于研究像重编程这样的动态细胞过程. 优化的miniTurbo PDB方法能够在初级细胞中快速进行蛋白质组分析,从而推进细胞命运研究.

关键词:
生物ID BioID 是一个生物ID.生物素是生物素中的一种.纤维母细胞细胞是如何形成的林氏病毒 (Lentivirus) 是一种病毒.质谱测量质量谱测量蛋白质与蛋白质的相互作用蛋白质组学是指蛋白质组学.靠近标签的标签.靠近依赖的生物化.重编程 重编程 是一种重编程.在Streptavidin中使用.

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科学领域:

  • 细胞生物学 细胞生物学
  • 蛋白质组学是指蛋白质组学.
  • 生物化学 生物化学

背景情况:

  • 在重编程和分化过程中了解细胞重组是阐明形态和命运变化的关键.
  • 靠近生物化 (PDB) 方法比传统的生物化学净化为亚细胞区的蛋白质特性提供了优势.
  • 像Bira* (BioID) 这样的第一代PDB酶通常需要很长的标签时间 (8-24小时).

研究的目的:

  • 在初级细胞中定义和优化时间解析的PDB协议.
  • 在小鼠纤维细胞模型中应用优化的PDB技术来研究体细胞重编程.
  • 为 PDB 适应其他动态细胞过程提供参考协议.

主要方法:

  • 使用高活性PDB酶,如miniTurbo,用于快速生物化信号 (分钟).
  • 开发并使用一种可诱导的lentiviral工具包用于BioID应用.
  • 在小鼠纤维细胞模型的体细胞重编程中,优化和应用lentivirally交付的miniTurbo构造.

主要成果:

  • 通过使用miniTurbo构造成功优化时间解析的PDB协议.
  • 展示了这些协议的应用,以研究体细胞重编程的动态过程.
  • 为研究人员在动态细胞研究中使用PDB建立了基线参考.

结论:

  • 时间解析的PDB,特别是在miniTurbo,可以对动态细胞事件进行高效的蛋白质组分析.
  • 开发的协议有助于研究体细胞重编程,并可适应其他细胞过程.
  • 这项工作促进了对细胞命运变化的机械基础的理解.