Ramachandran Shyam, Clarke Luka A, Scheetz Todd E, Amaral Margarida D, McCray Paul B
Interdisciplinary Program in Genetics, Department of Pediatrics, University of Iowa, Iowa City, IA, USA.
Methods Mol Biol. 2011;742:193-212. doi: 10.1007/978-1-61779-120-8_12.
To understand the links between CFTR mutations and the development of cystic fibrosis (CF) phenotypes, it is imperative to study the transcriptome in affected cell types. Microarray expression profiling provides a platform to study global gene expression in detail. This approach may provide the necessary information to segregate phenotypic characteristics of CF, differentiate between genetic or environmental factors, and assess the advent and progression of disease phenotypes. Moreover, if a "CF signature" of genes with altered expression is defined, this can be used to monitor effectiveness of treatment. We provide here detailed protocols and tips for collecting and preserving tissues and cells, and preparing total RNA. We also outline novel strategies for experimental design and data analysis, and describe some powerful gene and pathway discovery tools.
为了解囊性纤维化跨膜传导调节因子（CFTR）突变与囊性纤维化（CF）表型发展之间的联系，研究受累细胞类型的转录组至关重要。微阵列表达谱分析提供了一个详细研究全局基因表达的平台。这种方法可能提供必要信息，以区分CF的表型特征、区分遗传或环境因素，并评估疾病表型的出现和进展。此外，如果定义了表达改变的基因的“CF特征”，则可用于监测治疗效果。我们在此提供收集和保存组织与细胞以及制备总RNA的详细方案和提示。我们还概述了实验设计和数据分析的新策略，并描述了一些强大的基因和通路发现工具。
