Quick Guide For Adding A Reporter To Your cKO Model
Enhance your model with our exclusive TruView technology
Conditional knockout models enable more flexibility in experimental design compared with the older strategy of constitutive knockout. That flexibility is accompanied by a few additional requirements when planning how the line will be used, such as the extra element of a tissue-specific Cre line in breeding plans. Another step that must be considered is confirming that the knockout has happened as planned. To this end ingenious can create cKO models that incorporate reporter genes to label cells where recombination has occurred, providing unambiguous confirmation that the target allele has been inactivated. If this option could help you achieve your research goals please click here to access our short summary guide to different strategies for adding a reporter to your next cKO model.
Our guide to adding a reporter to a new cKO model describes the options and advantages of this approach. Adding a reporter can be particularly valuable when other means of validating knockout are difficult, for example in cases where antibodies against the protein of interest are difficult to use. Gene disruption using a cKO line can not be assumed, it should be verified for each experimental condition. Different Cre lines may interact with different cKO lines in unexpected ways that can’t be predicted in advance. This can lead to cases where, for example, a Cre line efficiently functions with one cKO line but not another, even in the same tissue. Different cKO lines each contain modifications in different genetic contexts, and subtle factors such as chromatin accessibility will affect Cre efficiency. Integrating a reporter directly into the cKO model cuts through all these challenges to give a readout of recombination at the level of individual cells. This can also make new experiments possible, particularly when using ingenious’ proprietary TruView technology. TruView incorporates a strong promoter for high levels of GFP expression in cells where knockout has occurred. In many cases the fluorescence from a TruView cKO model will be detectable in living cells.
If clearly marking cells where knockout has occurred would help with your next experiment click here to check out our guide. Then contact us to discuss your gene of interest with one of our technical specialists and find out how a reporter can enhance your next cKO model.