Investigation of the effects of Gabra3 gene knockout on sleep regulation in mouse neurons
10.16289/j.cnki.1002-0837.2025.03003
- VernacularTitle:Gabra3基因敲除对小鼠神经元细胞睡眠功能影响的探究
- Author:
Lizhong ZHANG
1
;
Yujie TANG
;
Yi SUN
;
Qi YU
;
Xuechun LU
Author Information
1. 山西医科大学基础医学院,太原 030000
- Keywords:
Gabra3;
slow-wave sleep;
CRISPR/Cas9;
multiomics;
hormone regulation
- From:Space Medicine & Medical Engineering
2025;36(3):206-213
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a Gabra3 gene knockout cell model and explore transcriptomic and proteomic alterations in murine neuronal cells,in order to investigate the molecular mechanisms underlying the increased depth of slow-wave sleep observed following Gabra3 deletion.Methods Multiple sgRNA sequences were designed,and the CRISPR/Cas9 system was used to knock out the Gabra3 gene in the murine GT1-7 neuronal cell line.Gene sequencing was performed to assess knockout efficiency,and TA cloning was used to validate the knockout results.Protein immunoblotting experiments were conducted to confirm the knockout,while cell proliferation assays were used to validate the knockout phenotype.Total RNA and protein were extracted for transcriptomic and proteomic sequencing,respectively.A range of bioinformatics analyses was conducted to assess the functional consequences of Gabra3 knockout in GT1-7 neuronal cells.Results Following Gabra3 gene knockout,pathways related to cortisol and aldosterone synthesis and secretion,as well as circadian rhythm,were significantly enriched.Three key genes,BMP2,GLI2,and DLL1,were identified.Proteomic profiling revealed widespread disturbances in protein expression following Gabra3 knockout.Conclusion Gabra3 gene knockout may increase slow-wave sleep depth by modulating the expression of hormone secretion-related genes and altering circadian regulatory pathways.
