Objective To construct a forebrain excitatory neuron-specific NR4A1 knockout mouse model based on the LoxP-Cre recombinase system and to investigate the effects of this knockout on cognitive function and anxiety-like behaviors in mice. Methods NR4A1 ^{flox / +} mice were self-crossed and screened to obtain NR4A1 ^{flox / flox} mice, which were then crossed with Camk2a-Cre mice. NR4A1 ^{flox / +}-Camk2a-Cre F1 mice were obtained and further self-crossed. The result ing offspring were genotyped using tail genomic DNA to screen out NR4A1 ^{flox / flox}-Camk2a-Cre mice, which were defined as NR4A1 conditional knockout mice, specifically in forebrain excitatory neurons. Cognitive ability was assessed using behavioral tests, including novel object recognition and Y-maze tests, and anxiety-like behaviors were evaluated using the elevated plus maze, open field, and light-dark box tests. Expression levels of NR4A1 in the hippocampus, as a key forebrain region, were detected by Western Blot and immunofluorescence. Results ( 1 ) Genotyping confirmed that NR4A1 ^{flox / flox} -Camk2a-Cre forebrain excitatory neuron-specific NR4A1 knockout mice were successfully obtained. (2) NR4A1 protein levels were significantly reduced in the hippocampus in knockout mice compared with the findings in control mice, as shown by Western Blot. Also, the intensity of NR4A1 immunofluorescent signals in the hippocampus was also lower in knockout group compared with that in control group mice. (3) In cognitive behavioral tests, there was no significant difference in novel object recognition index, number of entries, distance traveled, or residence time in the novel arm of the Y-maze between knockout group and control group mice. (4) In anxiety-like behavior tests, knockout group mice spent more time in the central area of the open field than control group mice, but there was no significant difference in the distance traveled or number of entries into the central area. In the light-dark box test, knockout group mice spent longer in the light compartment than control group mice, but there was no significant difference in the distance traveled or number of entries into the light compartment between the two groups. There was no significant difference between knockout group and control group mice in the elevated plus maze test. Conclusions This study successfully constructed a forebrain excitatory neuronspecific NR4A1 knockout mouse model. Knockout mice showed no significant changes in cognitive level but a significant reduction in anxiety levels. This model provides an important experimental tool for in-depth investigations of the role of the NR4A1 gene in the physiological functions and pathological mechanisms of forebrain regions.