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Calcium fiber photometry was used to reveal the role of KCC2 in the restoration of impaired GABAergic inhibition.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>CRISPRa system effectively upregulated KCC2 expression both in in\u2009vitro cell culture and in\u2009vivo brain region. Delivery of CRISPRa with adeno\u2010associated viruses resulted in upregulating the subicular KCC2 level, contributing to alleviating the severity of hippocampal seizure and facilitating the anti\u2010seizure effect of diazepam in a hippocampal kindling model. In a kainic acid\u2010induced epilepticus status model, KCC2 upregulation greatly increased the termination percentage of diazepam\u2010resistant epilepticus status with the broadened therapeutic window. More importantly, KCC2 upregulation attenuated valproate\u2010resistant spontaneous seizure in a kainic acid\u2010induced chronic epilepsy model. Finally, calcium fiber photometry showed CRISPRa\u2010mediated KCC2 upregulation partially restored the impaired GABA<jats:sub>A<\/jats:sub>\u2010mediated inhibition in epilepsy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Interpretation<\/jats:title><jats:p>These results showed the translational potential of adeno\u2010associated viruses\u2010mediated delivery of CRISPRa for treating neurological disorders by modulating abnormal gene expression that is directly associated with neuronal excitability, validating KCC2 as a promising therapeutic target for treating drug\u2010resistant epilepsy. 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