Objective The aim of this study was to establish a mouse model of H7N9 avian influenza A virus infection. Methods Seventy SPF healthy female BALB/c mice were used in this study. A/Anhui/1/2013 (H7N9) avian influenza virus was administered by intranasal instillation to BALB/c mice, inoculated in a dose of 50 μL 1×10⁸, 1×10⁷ or 1×10⁶ TCID₅₀, respectively, 10 mice in each group. Other 30 mice were used for virus titration and pathological examination. Ten mice were given saline as control group. The changes of body weight, clinical signs and death of the mice were observed every 24 h from 1 to 14 d. Blood and several organ samples were taken for pathological examination, and avian influenza A (H7N9) virus was detected with virus titration and immunohistochemistery (IHC). Results The mice developed typical clinical signs including body weight loss, ruffled fur and humped back. The peak of virus shedding from respiratory tract was observed on 2 days post inoculation (d.p.i.), and histopathological examination observed interstitial pneumonia. The virus was also detected in the brain, liver, spleen, kidney and intestine from inoculated mice. The inoculation of H7N9 virus elicited seroconversion titers up to 160. There was reduction of lymphocytes and increase of neutrophils in the blood. Conclusions The mouse model of H7N9 avian influenza virus infection established in this study show similar signs of human avian influenza. Therefore, it provides a useful working basis for research of the pathogenesis, drug development, and vaccine evaluation of this disease.