Takashi Nakamura, Sayako Honda, Shinichirou Ito, Toshihide Mizoguchi, Takehiro Yamamoto, Masataka Kasahara, Yasuaki Kabe, Koichi Matsuo, Makoto Suematsu
Journal of bone and mineral metabolism 2023 JulThe conditional manipulation of genes using the Cre recombinase-locus of crossover in P1 (Cre/loxP) system is an important tool for revealing gene functions and cell lineages in vivo. The outcome of this method is dependent on the performance of Cre-driver mouse strains. In most cases, Cre knock-in mice show better specificity than randomly inserted Cre transgenic mice. However, following knock-in, the expression of the original gene replaced by Cre is lost. We generated a new differentiated osteoblast- and osteocyte-specific Cre knock-in mouse line that carries the viral T2A sequence encoding a 2A self-cleaving peptide at the end of the coding region of the dentin matrix protein 1 (Dmp1) gene accompanied by the Cre gene. We confirmed that Dmp1-T2A-Cre mice showed high Cre expression in osteoblasts, osteocytes, odontoblasts, and periodontal ligament cells and that the 2A self-cleaving peptide efficiently produced both Dmp1 and Cre proteins. Furthermore, unlike the Dmp1 knockout mice, homozygous Dmp1-T2A-Cre mice showed no skeletal abnormalities. Analysis using the Cre reporter strain confirmed differentiated osteoblast- and osteocyte-specific Cre-mediated recombination in the skeleton. Furthermore, recombination was also detected in some nuclei of skeletal muscle cells, spermatocytes, and intestinal cells. 2A-Cre functions effectively in vivo, and Dmp1-T2A-Cre knock-in mice are a useful tool for studying the functioning of various genes in hard tissues. © 2023. The Japanese Society Bone and Mineral Research.
Takashi Nakamura, Sayako Honda, Shinichirou Ito, Toshihide Mizoguchi, Takehiro Yamamoto, Masataka Kasahara, Yasuaki Kabe, Koichi Matsuo, Makoto Suematsu. Generation of bicistronic Dmp1-Cre knock-in mice using a self-cleaving 2A peptide. Journal of bone and mineral metabolism. 2023 Jul;41(4):470-480
PMID: 37036533
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