Rhinogobio nasutus is an endemic fish species native to the middle and upper reaches of the Yellow River in China, renowned for its high economic and nutritional value. However, due to overfishing, human activities, and environmental pollution, it is now critically endangered. This study presents the construction of a telomere-to-telomere (T2T) reference genome for R. nasutus by integrating PacBio HiFi reads, Oxford Nanopore Technologies and Hi-C data. The assembled genome has a total size of 953.38 Mb and is anchored to 25 autosomes. Multiple assessments confirmed the high quality of the genome in terms of mapping rate (99.77% and 99.85%), completeness (BUSCO: 99.31%), and accuracy (QV: 47.05 and 53.10). Functional annotation was achieved for 98.16% of the 24,677 protein-coding genes, with a BUSCO score of 99.81%. This work not only facilitates the genetic conservation of R. nasutus but also provides a valuable resource for related evolutionary studies.