Cellular metabolism is foundational to all cellular activities. Targeting metabolic enzymes represents an effective approach to suppress pathogenic cell behavior and recently has received increasing attention in the fields of immunology and cancer. However, many of the metabolic pathways are essentially required by the majority of , if not all, the cells in the body, which greatly limits the clinical translation of metabolic targeting due to its broad toxicity. In this study, we show that CRISPR-mediated targeting of glycolysis in T cells in mice results in global loss of Th17 cells, whereas deficiency of the glycolytic enzyme Gpi1 selectively eliminates inflammatory encephalitogenic and colitogenic Th17 cells, without substantially affecting homeostatic microbiota-specific Th17 cells. Unlike glycolysis genes such as Gapdh, whose deficiency completely blocks glycolytic flux, Gpi1 encodes a functionally redundant enzyme, whose inactivation still supports adequate production of biosynthetic precursors and ATP in the homeostatic Th17 cells through enhanced compensatory activities of the pentose phosphate pathway and mitochondria respiration. In contrast, inflammatory Th17 cells experience a hypoxic microenvironment known to limit mitochondrial respiration, which is incompatible with loss of Gpi1. Our study suggests that inhibiting glycolysis by targeting Gpi1 could be an effective therapeutic strategy with minimum toxicity for Th17-mediated autoimmune diseases. Moreover, it demonstrates the remarkable plasticity of the metabolic network due to the redundant components which can be regulated by environmental factors, and, more importantly, that metabolic redundancies can be exploited for selective targeting of disease processes.