Genome Toxicity and Impaired Stem Cell Function after Conditional Activation of CreER^T2 in the Intestine.

With the tamoxifen-inducible CreER^T2 system, genetic recombination can be temporally controlled in a cell-type-specific manner in intact animals, permitting dissection of the molecular underpinnings of mammalian physiology. Here we present a significant drawback to CreER^T2 technology for analysis of intestinal stem cells. Using the intestine-specific Villin-CreER^T2 mouse strain, we observed delayed intestinal regeneration post irradiation. Villin-CreER^T2 activation was associated with DNA damage and cryptic loxP site cleavage. Analysis of stem cell-specific CreER^T2 strains showed that the genome toxicity impairs function of crypt base columnar stem cells, resulting in loss of organoid initiating activity. Importantly, the stem cell impairment is short-lived, with return to normal by 7 days post tamoxifen treatment. Our findings demonstrate that mouse genetic experiments that utilize CreER^T2 should consider the confounding effects of enhanced stem cell sensitivity to genome toxicity resulting from CreER^T2 activation.

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