Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments

Abstract Conditional creER-mediated gene inactivation or gene induction has emerged as a robust tool for studying gene functions in mouse models of tissue development, homeostasis, and regeneration. Here, we present a method to conditionally induce cre recombination in the mouse calvarial bone while...

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Autores principales:	Jue Hou, Charles P. Lin, Giuseppe Intini
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/72a0606ac7bd49b29fc018e3aa5a519a
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spelling	oai:doaj.org-article:72a0606ac7bd49b29fc018e3aa5a519a2021-12-02T18:03:26ZActivation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments10.1038/s41598-021-87611-22045-2322https://doaj.org/article/72a0606ac7bd49b29fc018e3aa5a519a2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87611-2https://doaj.org/toc/2045-2322Abstract Conditional creER-mediated gene inactivation or gene induction has emerged as a robust tool for studying gene functions in mouse models of tissue development, homeostasis, and regeneration. Here, we present a method to conditionally induce cre recombination in the mouse calvarial bone while avoiding systemic recombination in distal bones. To test our method, we utilized Prx1creER-egfp;td-Tomato mice and delivered 4-hydroxytamoxifen (4-OHT) to the mouse calvaria, subperiosteally. First, we showed that two calvaria subperiosteal injections of 10 µg of 4-OHT (3.3 mg of 4-OHT/kg of body weight) can induce local recombination as efficiently as two intraperitoneal systemic injections of 200 μg of tamoxifen (70 mg of tamoxifen/kg of body weight). Then, we studied the recombination efficiency of various subperiosteal calvaria dosages and found that two subperiosteal injections of 5 µg 4-OHT (1.65 mg of 4-OHT/kg of body weight) uphold the same recombination efficiency observed with higher dosages. Importantly, the result indicated that the low dosage does not induce significant systemic recombination in remote skeletal tissues. With the proposed local low dosage protocol, the recombination efficiency at the injection site (calvarial bone) reached 94%, while the recombination efficiency at the mandible and the digits was as low as the efficiency measured in control animals.Jue HouCharles P. LinGiuseppe IntiniNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jue Hou Charles P. Lin Giuseppe Intini Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
description	Abstract Conditional creER-mediated gene inactivation or gene induction has emerged as a robust tool for studying gene functions in mouse models of tissue development, homeostasis, and regeneration. Here, we present a method to conditionally induce cre recombination in the mouse calvarial bone while avoiding systemic recombination in distal bones. To test our method, we utilized Prx1creER-egfp;td-Tomato mice and delivered 4-hydroxytamoxifen (4-OHT) to the mouse calvaria, subperiosteally. First, we showed that two calvaria subperiosteal injections of 10 µg of 4-OHT (3.3 mg of 4-OHT/kg of body weight) can induce local recombination as efficiently as two intraperitoneal systemic injections of 200 μg of tamoxifen (70 mg of tamoxifen/kg of body weight). Then, we studied the recombination efficiency of various subperiosteal calvaria dosages and found that two subperiosteal injections of 5 µg 4-OHT (1.65 mg of 4-OHT/kg of body weight) uphold the same recombination efficiency observed with higher dosages. Importantly, the result indicated that the low dosage does not induce significant systemic recombination in remote skeletal tissues. With the proposed local low dosage protocol, the recombination efficiency at the injection site (calvarial bone) reached 94%, while the recombination efficiency at the mandible and the digits was as low as the efficiency measured in control animals.
format	article
author	Jue Hou Charles P. Lin Giuseppe Intini
author_facet	Jue Hou Charles P. Lin Giuseppe Intini
author_sort	Jue Hou
title	Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
title_short	Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
title_full	Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
title_fullStr	Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
title_full_unstemmed	Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
title_sort	activation of creer recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/72a0606ac7bd49b29fc018e3aa5a519a
work_keys_str_mv	AT juehou activationofcreerrecombinaseinthemousecalvariainduceslocalrecombinationwithouteffectsondistantskeletalsegments AT charlesplin activationofcreerrecombinaseinthemousecalvariainduceslocalrecombinationwithouteffectsondistantskeletalsegments AT giuseppeintini activationofcreerrecombinaseinthemousecalvariainduceslocalrecombinationwithouteffectsondistantskeletalsegments
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Activation of creER recombinase in the mouse calvaria induces local recombination without effects on distant skeletal segments

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