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Research Techniques Made Simple: Gene Knockout in Mice

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?.

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Research Techniques Made Simple: Gene Knockout in Mice

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  1. Research Techniques Made Simple:Gene Knockout in Mice Christian Günschmann, Elena Chiticariu, Bhavuk Garg, Merve Meliha Hiz, Yora Mostmans, Maria Wehner, Lukas Scharfenberger

  2. 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

  3. Tet-off System

  4. Tet-on System

  5. Tamoxifen-Inducible Cre-loxP System

  6. Confirmation of Tamoxifen-Mediated Knockout Concept of fibroblast-specific inducible TβRII knockout Loss of TβRII protein confirmed by western blot and IF wt Genotyping PCR Null Denton et al., J Invest Dermatol 129:194-204, 2009

  7. 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 Advantages and Limitations of Inducible Mice

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