The aim of the current study was to describe a novel approach of urethral reconstruction through minimally invasive harvesting of the bladder graft via endoscopic sub-mucosal dissection of water-jet. The records of two patients were reviewed, who underwent transurethral endoscopic surgical bladder mucosa graft harvest by water-jet and urethral reconstruction with informed consent. Case 1 was a 35-year-old male with anterior urethral stricture; case 2 was a 22-year-old male with secondary anterior urethral stricture and hypospadias following a failed hypospadias surgery. The two male patients successfully underwent urethral reconstruction using bladder mucosa graft harvested via endoscopic assisted by water-jet; no perforation, cysthemorrhagia or any other postoperative bladder-related complication was observed. Voiding cystourethrogram of case 1 indicated that the reconstructed urethra was unobstructed, and no recrudescence was observed within 4 months of follow-up. In case 2, dysuria had disappeared completely within 1 month of follow-up, and the urethra plate was successfully reconstructed by first-stage. To the best of our knowledge, this is the first report to demonstrate urethral reconstruction using a bladder mucosa graft harvested by transurethral endoscopic sub-mucosal dissection, assisted by water-jet. Transurethral endoscopic surgery may provide a minimally invasive approach instead of the traditional open surgery for harvesting bladder mucosa graft. Urethral reconstruction conducted with bladder mucosa graft harvested via endoscopic sub-mucosal dissection assisted by water-jet is a feasible and safe method, and the short-term follow-up results are encouraging.