Research Techniques Made Simple: Gene Knockout in Mice
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Research Techniques Made Simple:Gene Knockout in Mice
Christian Günschmann, Elena Chiticariu, Bhavuk Garg, Merve Meliha Hiz, Yora
Mostmans, Maria Wehner, Lukas Scharfenberger
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What Is Inducible Mouse Technology?
Allows the tissue/cell type-specific regulation of gene expression at a time point that is determined by the investigator – spatiotemporal control of gene knockout or gene knockin
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Tet-off System
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Tet-on System
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Tamoxifen-Inducible Cre-loxP System
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Confirmation of Tamoxifen-Mediated Knockout
Concept of fibroblast-specificinducible TβRII knockout
Genotyping PCR
Denton et al., J Invest Dermatol 129:194-204, 2009
Loss of TβRII protein confirmed by western blot and IF
wt
Null
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Advantages and Limitations of Inducible Mice
Advantages:
•Inducible mouse technology enables the investigator to determine where
and when to turn on or off genes of interest
•Allows assessment of the functional importance of overexpression or loss of
proteins in normal skin homeostasis and at different stages of disease
Limitations:
•Long-term treatment with the inducer can lead to side effects
•The system can be “leaky,” resulting in undesired expression of the gene of
interest