Carbon dioxide is increasingly used in insufflation during colonoscopy in adult patients; however, air insufflation remains the primary practice among pediatric gastroenterologists. This systematic review and meta-analysis aims to evaluate insufflation using CO2 versus air in colonoscopies in pediatric patients. Individualized search strategies were performed using MEDLINE, Cochrane Library, EMBASE, and LILACS databases following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines and Cochrane working methodology. Randomized control trials (RCTs) were selected for the present meta-analysis. Pooled proportions were calculated for outcomes including procedure time and abdominal pain immediately and 24 hours post-procedure. The initial search yielded 644 records, of which five RCTs with a total of 358 patients (CO2: n=178 versus air: n=180) were included in the final analysis. The procedure time was not different between the CO2 and air insufflation groups (mean difference, 10.84; 95% confidence interval CI, -2.55 to 24.22; p=0.11). Abdominal pain immediately post-procedure was significantly lower in the CO2 group (risk difference RD, -0.15; 95% CI; -0.26 to -0.03; p=0.01) while abdominal pain at 24 hours post-procedure was similar (RD, -0.05; 95% CI; -0.11 to 0.01; p=0.11). Based on this systematic review and meta-analysis of RCT data, CO2 insufflation reduced abdominal pain immediately following the procedure, while pain was similar at 24 hours post-procedure. These results suggest that CO2 is a preferred insufflation technique when performing colonoscopy in pediatric patients.