果蝇中单个信使核糖核酸分子的检测
Single mRNA Molecule Detection in Drosophila.
作者信息
Little Shawn C, Gregor Thomas
机构信息
Department of Cell and Developmental Biology, University of Pennsylvania Perelman School of Medicine, 1112 Biomedical Research Bldg. II/III, 421 Curie Blvd., Philadelphia, PA, 19104, USA.
Department of Physics, Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
出版信息
Methods Mol Biol. 2018;1649:127-142. doi: 10.1007/978-1-4939-7213-5_8.
Abstract
Single molecule fluorescent in situ hybridization (smFISH) enables quantitative measurements of gene expression and mRNA localization. The technique is increasingly popular for analysis of cultured cells but is not widely applied to intact organisms. Here, we describe a method for labeling and detection of single mRNA molecules in whole embryos of the fruit fly Drosophila melanogaster. This method permits measurements of gene expression in absolute units, enabling new studies of transcriptional mechanisms underlying precision and reproducibility in cell specification.
摘要
单分子荧光原位杂交（smFISH）能够对基因表达和mRNA定位进行定量测量。该技术在培养细胞分析中越来越受欢迎，但尚未广泛应用于完整生物体。在此，我们描述了一种在果蝇黑腹果蝇的全胚胎中标记和检测单个mRNA分子的方法。这种方法允许以绝对单位测量基因表达，从而能够对细胞特化中精确性和可重复性背后的转录机制进行新的研究。
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