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A Phenotyping Regimen for Genetically Modified Mice Used to Study Genes Implicated in Human Diseases of Aging
Published on: July 14, 2016
Generation and identification of a conditional knockout allele for the PSMD11 gene in mice
Linlin Zhao1, Jinming Zhao1, Yingying Zhang1
1Institute of Medical Sciences, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, 250033, China.
Background:
Our previous study have shown that the PSMD11 protein was an important survival factor for cancer cells except for its key role in regulation of assembly and activity of the 26S proteasome. To further investigate the role of PSMD11 in carcinogenesis, we constructed a conditional exon 5 floxed allele of PSMD11 (PSMD11flx) in mice.
Results:
It was found that homozygous PSMD11 flx/flx mice showed normal and exhibited a normal life span and fertility, and showed roughly equivalent expression of PSMD11 in various tissues, suggesting that the floxed allele maintained the wild-type function. Cre recombinase could induce efficient knockout of the floxed PSMD11 allele both in vitro and in vivo. Mice with constitutive single allele deletion of PSMD11 derived from intercrossing between PSMD11flx/flx and CMV-Cre mice were all viable and fertile, and showed apparent growth retardation, suggesting that PSMD11 played a significant role in the development of mice pre- or postnatally. No whole-body PSMD11 deficient embryos (PSMD11-/-) were identified in E7.5-8.5 embryos in uteros, indicating that double allele knockout of PSMD11 leads to early embryonic lethality. To avoid embryonic lethality produced by whole-body PSMD11 deletion, we further developed conditional PSMD11 global knockout mice with genotype Flp;FSF-R26CAG - CreERT2/+; PSMD11 flx/flx, and demonstrated that PSMD11 could be depleted in a temporal and tissue-specific manner. Meanwhile, it was found that depletion of PSMD11 could induce massive apoptosis in MEFs.
Conclusions:
In summary, our data demonstrated that we have successfully generated a conditional knockout allele of PSMD11 in mice, and found that PSMD11 played a key role in early and postnatal development in mice, the PSMD11 flx/flx mice will be an invaluable tool to explore the functions of PSMD11 in development and diseases.
Insights
PSMD11 protein is vital for mouse development, with its absence causing embryonic lethality. This study generated conditional knockout mice to investigate PSMD11
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- Previous research identified PSMD11 protein as crucial for cancer cell survival and 26S proteasome regulation.
- The PSMD11 protein's role in carcinogenesis requires further investigation.
Purpose of the Study:
- To generate a conditional exon 5 floxed allele of PSMD11 (PSMD11flx) in mice.
- To investigate the function of PSMD11 in mouse development and carcinogenesis.
Main Methods:
- Construction of a conditional exon 5 floxed allele of PSMD11 (PSMD11flx) in mice.
- Cre-mediated deletion of the floxed PSMD11 allele in vitro and in vivo.
- Generation of constitutive single allele deletion mice and conditional global knockout mice.
- Temporal and tissue-specific depletion of PSMD11 using Flp;FSF-R26CAG-CreERT2/+; PSMD11flx/flx mice.
Main Results:
- Homozygous PSMD11flx/flx mice were viable, fertile, and showed normal PSMD11 expression, indicating the floxed allele maintained wild-type function.
- Constitutive single allele deletion of PSMD11 resulted in growth retardation, while double allele knockout led to early embryonic lethality.
- Conditional PSMD11 depletion induced massive apoptosis in mouse embryonic fibroblasts (MEFs).
Conclusions:
- Successfully generated a conditional knockout allele of PSMD11 in mice.
- PSMD11 plays a critical role in both early and postnatal mouse development.
- The generated PSMD11flx/flx mice are valuable tools for studying PSMD11 functions in development and disease.
