Direct hematological toxicity and illegitimate chromosomal recombination caused by the systemic activation of CreERT2

Atsuko Yoshioka Higashi, Tomokatsu Ikawa, Masamichi Muramatsu, Aris N. Economides, Akira Niwa, Tomohiko Okuda, Andrew J. Murphy, Jose Rojas, Toshio Heike, Tatsutoshi Nakahata, Hiroshi Kawamoto, Toru Kita, Motoko Yanagita

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113 Citations (Scopus)


The CreERT2 for conditional gene inactivation has become increasingly used in reverse mouse genetics, which enables temporal regulation of Cre activity using a mutant estrogen binding domain (ERT2) to keep Cre inactive until the administration of tamoxifen. In this study, we present the severe toxicity of ubiquitously expressed CreERT2 in adult mice and embryos. The toxicity of Cre recombinase or CreERT2 in vitro or in vivo organisms are still less sufficiently recognized considering the common use of Cre/loxP system, though the toxicity might compromise the phenotypic analysis of the gene of interest. We analyzed two independent lines in which CreERT2 is knocked-in into the Rosa26 locus (R26CreERT2 mice), and both lines showed thymus atrophy, severe anemia, and illegitimate chromosomal rearrangement in hematopoietic cells after the administration of tamoxifen, and demonstrated complete recovery of hematological toxicity in adult mice. In the hematopoietic tissues in R26CreERT2 mice, reduced proliferation and increased apoptosis was observed after the administration of tamoxifen. Flow cytometric analysis revealed that CreERT2 toxicity affected several hematopoietic lineages, and that immature cells in these lineages tend to be more sensitive to the toxicity. In vitro culturing of hematopoietic cells from these mice further demonstrated the direct toxicity of CreERT2 on growth and differentiation of hematopoietic cells. We further demonstrated the cleavage of the putative cryptic/pseudo loxP site in the genome after the activation of CreERT2 in vivo. We discussed how to avoid the misinterpretation of the experimental results from potential toxic effects due to the activated CreERT2.

Original languageEnglish
Pages (from-to)5633-5640
Number of pages8
JournalJournal of Immunology
Issue number9
Publication statusPublished - 01-05-2009

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology


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