Abstract Disclosure: J. Park: None. S. Kim: None. E. Joo: None. J. Lee: None. Purpose: We aimed to investigate the genetic causes of suspected genetic short stature in 37 patients using next-generation sequencing (NGS) and describe their phenotypes and various genetic spectra. Methods: We reviewed the medical records of 50 patients who underwent genetic testing using NGS for suspected genetic short stature from June 2019 to December 2022. Patients with short stature caused by non-genetic factors or common chromosomal abnormalities were excluded. Thirty-seven patients from 35 families were enrolled in this study. We administered one of three genetic tests (targeted panel sequencing using two different panels or whole exome sequencing) to patients according to their phenotypes. Results: Clinical and molecular diagnoses were confirmed in 15 of the 37 patients, and the overall diagnostic yield was 40.5 %. Fifteen pathogenic/likely pathogenic variants were identified in 13 genes (ACAN, ANKRD11, ARID1B, CEP152, COL10A1, COL1A2, EXT1, FGFR3, NIPBL, NRAS, PTPN11, SHOX, SLC16A2). The diagnostic rate was high in the patients with small for gestational age (7/11, 63.6 %). Conclusions: Genetic evaluation using NGS can be helpful in patients with suspected genetic short stature who have clinical and genetic heterogeneity. Further studies are needed to develop patient selection algorithms and panels containing growth-related genes. Keywords: Dwarfism, Growth Hormone, Next-Generation Sequencing Presentation: 6/2/2024