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Summary: We explored the diversity and community composition of bacteria along a vertical gradient in Lake Issyk Kul, Kyrgyzstan, one of the world's largest and deepest brackish lakes. We identified 4904 bacterial amplicon sequence variants based on the 16S rRNA gene analysis and determined significant changes in the composition, responding mainly to depth and salinity. A higher abundance of Proteobacteria and Bacteroidetes was observed in the surface waters and lake tributaries. Cyanobacteria were more abundant in the deep chlorophyll maximum from 28.5 to 128 m, while Planctomycetes and Chloroflexi were dominant in the deepest layers, from 128 to 600 m. According to our machine learning analyses, depth and temperature were the most critical environmental factors, with strong effects on Proteobacteria, Planctomycetes and Chloroflexi, while oxygen was associated with the variations in Cyanobacteria. We also observed that with increasing depth, the alpha diversity values increased. The dominance of Planctomycetes and Chloroflexi in the deepest layers can only be seen in a few lakes of the world. However, the lake is facing increasing anthropogenic and climatic pressure. There is an urgent need to understand better the ecological role and function of these unique deep‐water microbial communities. [ABSTRACT FROM AUTHOR]