To ensure that the observed mitochondrial function and cognitive deficits were not a result of off-target effects, we predicted the off-target possibility of targeting NDUFS8 coding sequence “GGACTACACGCTATGACATT” in AAV-shRNA using NCBI database (Table S9), which demonstrated the high conservation of the NDUFS8 gene in this sequence (E value = 1.5). The gene discussed is NDUFS8; the disease is Cognitive impairment.