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Simple Summary: Invertase genes are among the important genes responsible for carbon metabolism in plants, significantly contributing to plant development and stress responses. In this study, for the first time, we performed genome-wide analysis for Invertase gene family in potato, identified and conducted expression profiling in different tissues by RNA seq analysis and validated it by Q-PCR. We also performed invertase family genes expression profiling under drought, salt and heat stress to elucidate their involvement in stress responses. Findings of this study will be helpful for future functional and genetic studies not only in potato but also in other plants. The potato is one of the most important and valuable crops in terms of consumption worldwide. However, abiotic stressors are the critical delimiters for the growth and productivity of potato. Invertase genes play key roles in carbon metabolism, plant development, and responses to stress stimuli. Therefore, a comprehensive genome-wide identification, characterization and expression analysis of invertase genes was performed in the potato. The current study identified 19 invertase genes, randomly distributed throughout the potato genome. To further elucidate their evolutionary, functional and structural relationship within family and with other plant species, we performed sequence and phylogenetic analysis, which segregated invertase genes into two main groups based on their sequence homology. A total of 11 genes are included in acidic invertases and 8 genes are in neutral or alkaline invertases, elucidating their functional divergence. Tissue specific expression analyses (RNA sequencing and qRT-PCR) of different plant tissues showed differential expression pattern. Invertase genes have higher expression in flower, leaf, root and shoot tissues, while under abiotic stress conditions, the expression of the invertase gene is significantly upregulated. Results of this study revealed that vacuolar and cell wall destined invertases are mainly the functional member genes of the invertase family. This study provides comprehensive data and knowledge about StINV genes in Solanum tuberosum for future genetic and epigenetic studies. [ABSTRACT FROM AUTHOR]