Aim: The influence on acute skeletal muscle transcriptomics of neuromuscular electrical stimulation (NMES), as compared to established exercises, is poorly understood. We aimed to investigate the effects on global mRNA‐expression in the quadriceps muscle early after a single NMES‐session, compared to the effects of voluntary knee extension exercise (EX), and to explore the discomfort level. Methods: Global vastus lateralis muscle gene expression was assessed (RNA‐sequencing) in 30 healthy participants, before and 3 h after a 30‐min session of NMES and/or EX. The NMES‐treatment was applied using textile electrodes integrated in pants and set to 20% of each participant's pre‐tested MVC mean (±SD) 200 (±80) Nm. Discomfort was assessed using Visual Analogue Scale (VAS, 0–10). The EX‐protocol was performed at 80% of 1‐repetition‐maximum. Results: NMES at 20% of MVC resulted in VAS below 4 and induced 4448 differentially expressed genes (DEGs) with 80%‐overlap of the 2571 DEGs of EX. Genes well‐known to be up‐regulated following exercise, for example, PPARGC1A, ABRA, VEGFA, and GDNF, were also up‐regulated by NMES. Gene set enrichment analysis demonstrated many common pathways after EX and NMES. Also, some pathways were exclusive to either EX, for example, muscle tissue proliferation, or to NMES, for example, neurite outgrowth and connective tissue proliferation. Conclusion: A 30‐min NMES‐session at 20% of MVC with NMES‐pants, which can be applied with an acceptable level of discomfort, induces over 4000 DEGs, of which 80%‐overlap with DEGs of EX. NMES can induce exercise‐like molecular effects, that potentially can lead to health and performance benefits in individuals who are unable to perform resistance exercise. [ABSTRACT FROM AUTHOR]