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未分化胚胎细胞转录因子 1 的体内和体外动力学。

In Vivo and in vitro dynamics of undifferentiated embryonic cell transcription factor 1.

机构信息

Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA ; Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.

出版信息

Stem Cell Reports. 2014 Feb 20;2(3):245-52. doi: 10.1016/j.stemcr.2014.01.007. eCollection 2014 Mar 11.

Abstract

Pluripotent stem cells retain the ability to differentiate into the three germ layers and germline. As a result, there is a major interest in characterizing regulators that establish and maintain pluripotency. The network of transcription factors continues to expand in complexity, and one factor, undifferentiated embryonic cell transcription factor 1 (UTF1), has recently moved more into the limelight. To facilitate the study of UTF1, we report the generation and characterization of two reporter lines that enable efficient tracking, mapping, and purification of endogenous UTF1. In particular, we include a built-in biotinylation system in our targeted locus that allows efficient and reliable pulldown. We also use this reporter to show the dynamic regulation of Utf1 in distinct stem cell conditions and demonstrate its utility for reprogramming studies. The multipurpose design of the reporter lines enables many directions of future study and should lead to a better understanding of UTF1's diverse roles.

摘要

多能干细胞保留了分化为三个胚层和生殖系的能力。因此,人们对鉴定建立和维持多能性的调节因子非常感兴趣。转录因子网络的复杂性不断扩大,最近有一个因子,未分化胚胎细胞转录因子 1(UTF1),更加引人注目。为了便于研究 UTF1,我们报告了两种报告基因系的产生和特性,这两种报告基因系能够有效地跟踪、映射和纯化内源性 UTF1。特别是,我们在靶向基因座中内置了一个生物素化系统,允许高效可靠的下拉。我们还使用该报告基因来显示 Utf1 在不同干细胞条件下的动态调节,并证明其在重编程研究中的用途。报告基因系的多功能设计能够为未来的许多研究方向提供支持,应该有助于更好地理解 UTF1 的多种作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d7d4/3964277/d08552bb6a7f/gr1.jpg

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