Neuropathic pain, characterized by persistent or intermittent spontaneous pain as well as some unpleasant abnormal sensations, is one of the most prevalent health problems in the world. Ectopic nerve activity, central and peripheral nociceptive sensitization and many other potential mechanisms may participate in neuropathic pain. The complexity and ambiguity of neuropathic pain mechanisms result in difficulties in pain management, and existing treatment plans provide less-than-satisfactory relief. In recent years, single-cell RNA sequencing (scRNA-seq) has been increasingly applied and has become a powerful means for biological researchers to explore the complexity of neurobiology. This technique can be used to perform unbiased, high-throughput and high-resolution transcriptional analyses of neuropathic pain-associated cells, improving the understanding of neuropathic pain mechanisms and enabling individualized pain management. To date, scRNA-seq has been preliminarily used in neuropathic pain research for applications such as compiling a dorsal root ganglion (DRG) atlas, identifying new cell types and discovering gene regulatory networks associated with neuropathic pain. Although scRNA-seq is a relatively new technique in the neuropathic pain field, there have been several studies based on animal models. However, because of the various differences between animals and humans, more attention should be given to translational medicine research. With the aid of scRNA-seq, researchers can further explore the mechanism of neuropathic pain to improve the clinical understanding of the diagnosis, treatment and management of neuropathic pain.