High Resolution Genome Wide Expression Analysis of Single Myofibers Using SMART-Seq

Skeletal muscle is a heterogeneous tissue. Individual myofibers that make up the contractile muscle tissue exhibit variation in their metabolic and contractile properties. Although there are biochemical and histological assays to study myofiber heterogeneity, methods to analyze the transcriptomes of individual myofibers are lacking. We have developed single myofiber RNA-Seq (smfRNA-Seq) to analyze the whole transcriptome of an individual myofiber by combining single fiber isolation with Switching Mechanisms at 5' end of RNA Template (SMART) technology. Our method provides high-resolution genome wide expression profiles of single myofibers. This method will be useful to study developmental and age-related dynamics in the composition of skeletal muscle. Gene expression analysis of single myofibers, groups of 5 or 20 myofibers, and whole muscle, each in triplicate骨骼肌是一类异质性组织。构成收缩性肌肉组织的单个肌纤维，其代谢与收缩特性存在差异。尽管已有生化和组织学检测手段可用于研究肌纤维异质性，但目前仍缺乏能够分析单个肌纤维转录组的方法。本研究开发了单肌纤维RNA测序（single myofiber RNA-Seq, smfRNA-Seq）技术，通过将单纤维分离与RNA模板5'端转换机制（Switching Mechanisms at 5' end of RNA Template, SMART）技术相结合，实现对单个肌纤维的全转录组分析。该方法可获取单个肌纤维的高分辨率全基因组表达谱，能够用于研究骨骼肌组成的发育及年龄相关动态变化。本数据集包含单个肌纤维、5个或20个肌纤维群组以及全肌肉组织的基因表达分析数据，且每组样本均设置三次生物学重复。