在多能干细胞模型中的单细胞转录扰动组
Elisa Balmas1, Maria L Ratto2, Kirsten E Snijders2
1Molecular Biotechnology Center "Guido Tarone", University of Turin, Turin, Italy. elisa.balmas@unito.it.
Molecular systems biology
|December 10, 2025
概括
我们创建了iPS2-seq,这是人类诱导多能干细胞 (hiPSCs) 的新型查平台. 这种工具可以在有机体等复杂模型中精确分析基因功能,从而推进功能基因组学研究.
科学领域:
- 干细胞生物学 干细胞生物学
- 功能性基因组学 功能性基因组学
- 发展生物学 发展生物学
背景情况:
- 在人类诱导多能干细胞 (hiPSCs) 中的功能基因组学选至关重要,但在技术上具有挑战性.
- 现有的方法很难区分真正的扰动效应和固有的细胞变异性.
- 复杂的hiPSC衍生模型,如有机体,对遗传查构成独特的障碍.
研究的目的:
- 开发一个可诱导,克隆意识的选平台,用于hiPSC衍生品.
- 为了使功能丧失效应的表型不可知,单细胞分辨率.
- 在基于hiPSC的模型中促进严格的功能基因组学,包括有机体.
主要方法:
- 开发iPS2-seq,一个可诱导和克隆意识的查平台.
- 与微流体或分池单细胞RNA测序进行集成,用于多原子分析.
- 使用专用分析管道,catcheR,以简化实验设计和数据解释.
主要成果:
- iPS2-seq成功地将真正的遗传干扰效应与表观遗传和遗传变异性区分开来.
- 已证明在单层心肌细胞和有机体中适用于针对先天性心脏病基因的应用.
- 确定了表观遗传神经切皮原始化作为胚胎层分化的偏差,并揭示了SMAD2在心脏前代特征中的作用.
结论:
- iPS2-seq为hiPSC衍生物和有机体中的高通量功能基因组学提供了一个强大的平台.
- 该平台克服了以前方法的局限性,使得基因功能的精确剖析成为可能.
- 这项工作为研究人类发育和疾病使用先进的干细胞模型打开了新的可能性.
相关概念视频
01:32Chromatin Modification in iPS Cells
2.1K
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.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
2.1K
01:29Somatic to iPS Cell Reprogramming
2.6K
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...
2.6K
01:13Induced Pluripotent Stem Cells
27.2K
Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
27.2K
01:06Induced Pluripotent Stem Cells
5.3K
Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic...
Somatic...
5.3K
02:33Combinatorial Gene Control
9.4K
Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
9.4K
01:14Maintenance of the ES Cell State
2.6K
The cells of the blastocyst inner cell mass only remain pluripotent for a short time. This state of pluripotency and self-renewal can be maintained in embryonic stem (ES) cell culture by adding specific chemicals or growth factors to ensure the cells can continue dividing and later differentiate into different cell types. In some cases, the cells are grown on a feeder layer of differentiated cells, which provides the growth factors and extracellular matrix components necessary for stem cell...
2.6K