Genomics plays a crucial role in healthcare, enhancing diagnostics, risk assessments, and therapeutic interventions. However, many healthcare professionals, including nurses, face challenges when it comes to integrating genomics into their practice. This study aims to evaluate the genomic knowledge of nurse practitioners in Greece by translating and validating the Genomic Nursing Concept Inventory (GNCI). A cross-sectional study involving 324 nurses was conducted in Crete, Greece. The GNCI, a 31-item questionnaire, was translated and validated for the Greek context to assess nurses' genomics competence. The Greek version of the GNCI demonstrated acceptable reliability (Cronbach's alpha = 0.622). The confirmatory factor analysis indicated a satisfactory fit for the one-factor model. However, the mean GNCI score revealed significant knowledge gaps, with nurses answering only 30.1% of questions correctly. Notably, nurses showed a better understanding of genomic basics and mutations compared to inheritance and genomic healthcare applications. The present study successfully translated and validated the GNCI in Greek and highlighted important genomic-related knowledge gaps among Greek nurses, emphasizing the urgent need for more targeted educational interventions. By enhancing genomic literacy through curriculum integration and professional development, nurses could be better prepared, leading to improved patient care and healthcare outcomes in Greece.