To describe the safety and feasibility of a running urethrovesical anastomosis (UVA) in robot-assisted radical prostatectomy (RARP) using a unidirectional self-locking barbed suture. Fifty-one consecutive patients with organ-confined prostate cancer underwent RARP by one of two experienced surgeons. UVA was performed in two layers, using a unidirectional barbed suture fashioned into a double-ended stitch. Perioperative outcomes and 30-day complications were recorded. All anastomoses were performed without assistance and without tying a knot. Median time for entire dual-layer anastomosis was 14.0 minutes (interquartile range [IQR]: 12-20) and that for urethrovesical anastomosis was 11 minutes (IQR: 9-15). Not having to rely on an assistant to follow the suture decreased instrument clashes, entangling of the suture around an instrument, and made the anastomosis faster. Eight patients underwent anterior/lateral reconstruction of the bladder neck, and there were no leaks on cystography at 1 week. We describe the first reported clinical experience with a novel technique of performing UVA during RARP that is safe and efficient. Using the barbed wound closure device prevents slippage, precluding the need for assistance, knot tying, and constant reassessing of anastomosis integrity.