Crop genetic resources are crucial for sustaining human life, agricultural productivity, and diversity at different levels. Ethiopia is a country rich in biodiversity, which puts the country among the major world seats for crop origin and diversity. Due to factors like land use change, climate change and the introduction and expansion of modern varieties, crop genetic resources face genetic erosion. In light of this fact, the Ethiopian Gene Bank was established in 1976. The Ethiopian Biodiversity Institute (EBI) hosts the National Gene Bank and is mandated to conserve the biodiversity wealth including traditional knowledge while ensuring fair and equitable sharing of benefits arising from the resources. Collection missions are conducted based on factors such as the crop’s economic importance and threat level. The Ethiopian Gene Bank (seed storage facility) holds over 85,000 accessions of 201 crop species. As crops with recalcitrant seeds cannot be kept in cold storage, over 7500 accessions of various crop species are conserved in five field gene banks. Over 35 crop species are conserved across 42 community seed banks operating in the country. Cognizant of the importance, the Ethiopian Biodiversity Institute is carrying out the characterization of conserved accessions. Characterization of EBI’s collection has revealed the existence of substantial variation in the collection and the potential of the resource for crop improvement. This chapter presents the details of the activities carried out in the Ethiopian Gene Bank with regard to the conservation and sustainable utilization of crop genetic resources.