Research Techniques Made Simple:Gene Knockout in Mice

Christian Günschmann, Elena Chiticariu, Bhavuk Garg, Merve Meliha Hiz, Yora

Mostmans, Maria Wehner, Lukas Scharfenberger

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

Tet-off System

Tet-on System

Tamoxifen-Inducible Cre-loxP System

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

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