The study of micro RNA (miRNA) expression and function, a largely unexplored area of human muscle biology, may provide novel data regarding the development of targeted approaches that optimize skeletal muscle responses to exercise and amino acid manipulations. miRNAs are ubiquitously expressed, small noncoding RNAs that modulate posttranscriptional gene expression. Quantifying miRNA expression and predicting function as regulators of both single targets and complex networks is technically challenging and requires a combined approach of bioinformatics, molecular, and systems biology. Recent evidence suggests that the expression of muscle-specific miRNAs (myomirs), including miR-1, miR-133a/b, miR-206, and miR-499, is modulated by essential amino acid ingestion, endurance exercise, and endurance exercise training. The expression of miRNAs has also been implicated in the anabolic intracellular signaling and muscle hypertrophic response associated with resistance exercise training. Although these findings are intriguing, comprehensive human trials assessing functional outcomes associated with changes in miRNA expression in response to exercise and nutrition interventions have not been conducted. This article reviews the current understanding of miRNA biology and includes analytical techniques used to detect miRNA expression and methods to predict function. The intent is to provide the framework for future research studies that use miRNA analysis in an effort to elucidate optimal exercise and nutritional countermeasures for the prevention of muscle loss.