Hello!
We are generating conditional knockout mice using CX3CR1 Cre for a particular gene. During this process, we are performing tail cuts for genomic extraction and genotyping. We've designed primers for flox PCR to detect a deletion band if Cre is present. However, we've encountered an issue. Despite the tail typically having fewer cells like those containing CX3CR1 Cre (such as microglia, macrophages, dendritic cells), deletion bands are occasionally detected in tail genotyping. In some mice, these bands appear stronger than the normal flox allele bands. Conversely, in other mice with Cre, no deletion bands are observed at all. Could this be due to low-level expression of CX3CR1 in some cells in the tail? Should germline depletion be considered in these scenarios?
Thanks,
Hi, Which mice do you have? If they don't have the additional tamoxifen-dependence then neurons also sometimes express the cre (probably early in development). Even the tamoxifen-dependent lines might have some 'leak'. You can find a lot of information if you have JAX lines on their website...Non-myeloid cell expression also might be dependent on the floxed construct i.e. some recombine more efficiently then others (or might even undergo spontaneous recombination with no cre at all).
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