在哺乳动物细胞中控制转录因子活性和骨质母细胞分化,使用一种进化的小分子依赖性整蛋白
Courtney M Yuen1, Stephen J Rodda, Steven A Vokes
1Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, Massachusetts 02138, USA.
Journal of the American Chemical Society
|July 6, 2006
概括
通过使用4-胺 (4-HT) 的工程整因,可以精确控制哺乳动物细胞中的蛋白质活性. 这个系统允许小分子诱导的基因特异性,剂量依赖的翻译后调节.
科学领域:
- 分子生物学分子生物学
- 合成生物学 合成生物学
- 生物化学 生物化学
背景情况:
- 蛋白质是催化拼接的蛋白质元素,切除自己并加入侧面的蛋白质.
- 之前的研究开发了依赖酵母中的4-胺 (4-HT) 的基于素的开关.
- 通过外部分子控制蛋白质活动对于生物研究和治疗至关重要.
研究的目的:
- 在哺乳动物细胞中证明进化型整蛋白对4-HT依赖蛋白拼接的有效性.
- 在不同细胞类型中描述该系统的动力学和剂量依赖性.
- 应用这个系统来控制基因功能和复杂的生物过程.
主要方法:
- 对于4-HT依赖的整数的定向演化.
- 在NIH3T3和HEK293细胞中因联体诱导的蛋白质拼接的表征.
- 进化整蛋白的应用,以控制刺途径介质 (Gli1,Gli3) 和骨质细胞分化.
主要成果:
- 进化后的整因在哺乳动物细胞中有效调解了4-HT-依赖蛋白质拼接.
- 该系统展示了通用性,速度和剂量依赖控制.
- 使用4-HT诱导性整蛋白系统实现了Gli1,Gli3和骨质母细胞分化的功能控制.
结论:
- 进化的小分子依赖性整蛋白为哺乳动物系统中蛋白质活性的精确,翻译后控制提供了多功能平台.
- 这项技术使得基因特异性,剂量依赖性和外部调节的生物途径和过程的操纵成为可能.
- 该系统有可能在基因疗法,药物发现和基本生物研究等领域应用.
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Transcription
Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
Transcription
Transcription is the synthesis of RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in correctly synthesizing messenger RNA (mRNA). Transcriptional regulation is responsible for the differentiation of different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds of RNA Molecules
In eukaryotes,...
Transcription Can Produce Different Kinds of RNA Molecules
In eukaryotes,...
General Transcription Factors
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
Somatic to iPS Cell Reprogramming
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 for this...
Methods of Nuclear Reprogramming
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 injury repair.
