Construction and phenotypic study of heterozygous knockout mice of tumor necrosis factor receptor associated factor 2
10.19405/j.cnki.issn1000-1492.2025.07.018
- Author:
Weikang Wang
1
;
Shujun Zuo
1
;
Jintao Gu
1
;
Fuyuan Guo
1
;
Haozhou Guo
1
;
Chenchen Han
1
;
Wei Wei
1
Author Information
1. Institute of Clinical Pharmacology，School of Pharmaceutical Sciences，Anhui Medical University， Key Laboratory of Anti Inflammatory and Immune Drugs of the Ministry of Education，Anhui Collaborative Innovation Center for Anti Inflammatory and Immune Drugs，Hefei 230032
- Publication Type:Journal Article
- Keywords:
TRAF2;
CRISPR/Cas9;
gene knockout;
genetically engineered mice;
genotyping;
phenotypic study
- From:
Acta Universitatis Medicinalis Anhui
2025;60(7):1291-1296
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To generate heterozygous TRAF2 knockout mice, the CRISPR/Cas9 technology was successfully employed. These mice were served as a valuable model to explore the pathological mechanisms underlying inflammatory and immune disorders mediated by abnormal TNF-α-TRAF2 signaling and to develop new therapeutic targets.
Methods : A vector targeting the knockout of the TRAF2 gene was constructed. Lead RNA and Cas9 Mrna were introduced into the fertilized eggs of C57BL/6JGpt mice through microinjection to mediate the TRAF2 gene mutation in mice. The mouse tail protein was extracted and the genotype of the F0 generation was determined by PCR and Western blot. TRAF2+/- mice were successfully obtained. F0 generation mice were backcrossed with C57BL/6JGpt wild-type mice to obtain stable TRAF2+/- mice for propagation and subsequent experiments. The body weight of TRAF2+/- mice was detected; Western blot was used to detect the expression of TRAF2 in the spleen, liver and kidney tissues of TRAF2+/- mice. The development of spleen, liver and kidney tissues in TRAF2+/- mice was detected by HE staining.
Results :PCR identification using specific primers demonstrated that TRAF2+/- mice exhibited a target band at 679 bp. Western blot analysis results indicated that, compared with the WT group, the expression of TRAF2 in the tail protein of TRAF2+/- mice was significantly reduced(P+/- mice had a lower body weight compared to their littermate WT mice(P+/- mice was decreased(P+/- mice and WT mice.
Conclusion:The successful construction of TRAF2+/- mice has provided an important animal model for exploring the role of TRAF2 in developmental regulation, revealing the mechanism of inflammatory immune diseases mediated by abnormal TNF-α-TRAF2 signaling, and screening related drug targets.
- Full text:2026030516180687790肿瘤坏死因子受体相关因子2杂合子敲除小鼠构建和表型研究_王伟康.pdf
