Conditional Knockout Model with Monitoring of the Gene Ablation Efficacy

Conditional Knockout models, either in a tissue-specific manner or in an inducible manner, are now routinely used models to study gene functions and validate targets. The conditional deletion effectiveness (in how many cells of the tissue of interest, both alleles are deleted) is a key parameter for the quality and reliability of the model. To monitor this efficacy and validate the reliability of the model, scientists are now using this technology to target a reporter gene in the locus of interest which is switched on when the deletion occurs. Measuring the reporter gene expression quantifies the level of deletion obtained in a tissue or at the cell level indicating which cell is knocked out and which one is not. By providing a means to validate the conditional Knockout efficacy, this technology enables the creation of reliable and valuable conditional Knockout models for target validation.

 

Case study: Tracking of Knockout cells in vivo when no good antibody is available.

Adapted from Schnütgen et al, Nat Biotechnol 2003, A directional strategy for monitoring Cre-mediated recombination at the cellular level in the mouse.

Monitoring of a Cre-mediated gene deletion

 

Efficient Monitoring of the Inactivated Gene

Monitoring a Cre-mediated deletion of a gene of interest (Retinoic acid Receptor gamma).

Before induction, no staining was detected (top).

After induction, reporter activity was detected in Rar gamma-expressing cells (bottom).