胚胎干细胞和囊胚中基因表达的聚合酶克隆分析。

Polony analysis of gene expression in ES cells and blastocysts.

作者信息

Rieger C, Poppino R, Sheridan R, Moley K, Mitra R, Gottlieb D

机构信息

Department of Anatomy and Neurobiology, Washington University School of Medicine, 660 S. Euclid Avenue, St Louis, MO 63110, USA.

出版信息

Nucleic Acids Res. 2007;35(22):e151. doi: 10.1093/nar/gkm1076. Epub 2007 Dec 10.

DOI:10.1093/nar/gkm1076

PMID:18073198

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2190707/

Abstract

Expression profiling of stem cells is challenging due to their small numbers and heterogeneity. The PCR colony (polony) approach has theoretical advantages as an assay for stem cells but has not been applied to small numbers of cells. An assay has been developed that is sensitive enough to detect mRNAs from small numbers of ES cells and from fractions of a single mouse blastocyst. Genes assayed include Oct3, Rex1, Nanog, Cdx2 and GLUT-1. The assay is highly sensitive so that multiple mRNAs from a single blastocyst were easily detected in the same assay. In its present version, the assay is an attractive alternative to conventional RT-PCR for profiling small populations of stem cells. The assay is also amenable to improvements that will increase its sensitivity and ability to analyze many cDNAs simultaneously.

摘要

由于干细胞数量少且具有异质性，对其进行表达谱分析具有挑战性。PCR集落（polony）方法作为一种干细胞检测方法具有理论优势，但尚未应用于少量细胞。现已开发出一种检测方法，其灵敏度足以检测少量胚胎干细胞以及单个小鼠囊胚各部分的mRNA。检测的基因包括Oct3、Rex1、Nanog、Cdx2和GLUT-1。该检测方法高度灵敏，因此在同一次检测中能够轻松检测到来自单个囊胚的多种mRNA。就目前的版本而言，该检测方法是对少量干细胞群体进行表达谱分析的传统逆转录聚合酶链反应（RT-PCR）的一种有吸引力的替代方法。该检测方法也易于改进，这将提高其灵敏度以及同时分析多个互补DNA（cDNA）的能力。