Functional characterization of de novo frameshift mutation in IRF8
10.3969/j.issn.1672-8467.2024.03.010
- VernacularTitle:IRF8新发移码突变的基因功能分析
- Author:
Le-Ying LI
1
;
Yao CHEN
;
Wei-Tao ZHOU
;
Chen HE
;
Duan-Wu ZHANG
;
Li-Ling QIAN
Author Information
1. 国家儿童医学中心/复旦大学附属儿科医院呼吸科 上海 201102;国家儿童医学中心/复旦大学附属儿科医院儿科研究所 上海 201102
- Keywords:
interferon regulatory factor 8(IRF8);
frameshift mutation;
recurrent infection;
transcription regulation
- From:
Fudan University Journal of Medical Sciences
2024;51(3):359-367
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study and verify the function of de novo interferon regulatory factor(IRF8)frameshift mutation detected in an etiology screening of the cohort of children with recurrent pneumonia at the molecular level.Methods The recombinant overexpression plasmids with wildtype or mutated IRF8 genes were constructed to transiently transfect HEK293T cells,or packed into lentivirus to infect two kinds of immune cell lines.Q-PCR,Western blot,immunofluorescence and other experimental assays were performed to explore the differences of expression and the regulatory effect on downstream genes associated with inflammation.Results The recombinant vectors with wildtype or mutated IRF8 genes were constructed successfully,and the efficiency of transfection by plasmids and infection by packed lentivirus was remarkable as well.Compared with wildtype,the molecular weight of IRF8 variant was slightly increased,while the expression level presents in opposition,even if on transcription level.Moreover,the localization of IRF8 variant was detected in abundance in nucleus rather than cytoplasm,and its inhibition effect was enhanced on the downstream ISRE element in comparison with the wildtype IRF8 protein.Conclusion The de novo frameshift mutation was presumed as gain-of-function(GOF)mutation.
