Objective: This PUBA study aimed to assess the efficacy of fractional CO2 laser in the treatment of genitourinary syndrome of menopause (GSM). Methods: GSM symptoms were assessed before, 1 month after the first session and 1 month after the third session of laser (3 sessions with a 30 days interval between them) in 60 women (median, interquartile range: 55, 49–69). Subjective (visual analog scale) and objective (Vaginal Health Index, VHIS; Vaginal Maturity Index/Frost Index; Spanish Overactive Bladder Questionnaire-Short Form, USMEX Spanish OAB-qSF and Female Sexual Function Index, FSFI) measures were used during the study period to assess CO2 fractionated laser treatment outcomes compared to baseline. Results: Fractional CO2 laser treatment was effective to improve GSM symptoms (vaginal dryness, vaginal itching, vaginal burning, dyspaurenia, dysuria, urinary urgency; P < 0.001) after three sessions, as well as VHIS (median, interquartile range: 13, 10–15 at baseline vs. 21, 20–23 at the fourth month follow up; P < 0.001), Frost Index (median, interquartile range: 28, 24–31 at baseline vs. 8, 6–10 at the fourth month follow up; P < 0.001), USMEX (median, interquartile range: 56, 46–68 at baseline vs 14, 13–16 at the fourth month follow up: P < 0,001) and FSFI (median, interquartile range: 5, 2–14 at baseline vs 30, 28–32). Conclusions: In this sample, the data suggests that fractionated CO2 laser is an effective alternative for GSM treatment with positive outcomes that persists over time. Lasers Surg. Med. 51:509–515, 2019. © 2019 Wiley Periodicals, Inc.