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相关概念视频

Production of Pharmaceuticals01:30

Production of Pharmaceuticals

Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
The Proteasome02:18

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
The Proteasome01:13

The Proteasome

Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3 (ubiquitin...
The Proteasome Structure01:17

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The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...

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相关实验视频

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Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
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一个基于蛋白酶的可编程蛋白质分泌平台,用于治疗应用.

Xinyi Wang1, Liping Kang1, Deqiang Kong1

  • 1Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.

Nature chemical biology
|October 23, 2023
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概括

本研究介绍了一种基于蛋白酶的快速蛋白质分泌系统 (PASS),用于细胞疗法. 通过能够快速,按需分泌治疗性蛋白质,推进精确医学.

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科学领域:

  • 生物医学科学 生物医学科学
  • 细胞治疗工程 细胞治疗工程
  • 分子生物学分子生物学

背景情况:

  • 目前的细胞疗法依赖于转录/翻译来获得治疗输出.
  • 现有的基因控制系统缺乏快速蛋白质分泌能力.
  • 工程细胞需要准确和及时的治疗蛋白质的输送.

研究的目的:

  • 为基于细胞的疗法开发一种基于蛋白酶的新型快速蛋白质分泌系统 (PASS).
  • 为了证明PASS变体在不同治疗环境中的灵活性和有效性.
  • 为了使预翻译蛋白质从内细胞网膜迅速,可诱导分泌.

主要方法:

  • 开发了三种PASS变种:chemPASS,antigenPASS和optoPASS. 这三种变种的研发过程包括:
  • 使用ER检索信号来保留预转换的蛋白质.
  • 使用可诱导蛋白酶来控制蛋白质释放.
  • 在糖尿病,癌症和疼痛的小鼠模型中进行体内验证.

主要成果:

  • chemPASS证明了快速的胰岛素分泌,可以逆转糖尿病小鼠的高血糖症.
  • 抗原PASS通过大酶B和穿孔素分泌诱导了向瘤细胞亡.
  • 在各种条件下,optoPASS在几分钟内使光诱导治疗性分泌成为可能.
  • 通过在各种疾病模型中促进了快速治疗蛋白质的输送.

结论:

  • 基于蛋白酶的快速蛋白质分泌系统 (PASS) 为基于细胞的疗法提供了一个灵活的平台.
  • 通过能够快速,可控的治疗蛋白质的交付,提高精确医学.
  • 这项技术有可能加速先进细胞疗法的开发和采用.