Conditional Ccnd1 Overexpression in Pax7+ Muscle Stem Cells

TRE-Ccnd1^WT and TRE-Ccnd1^K112E mice transgenic for human Ccnd1 with or without the K112E mutation under TRE regulation have been published previously^{31 (link),32 (link)}. Pax7^rtTA mice contain rtTA-M2 followed by an IRES, mouse Pax7, and polyadenylation tail knocked in to the first exon of the Pax7 locus. Both lines were maintained by breeding heterozygous animals to wild-type animals. Experimental animals (Pax7^rtTA/+, Pax7^rtTA/+; TRE-Ccnd1^WT, and Pax7^rtTA/+; TRE-Ccnd1^K112E) were generated by intercrossing the two lines and aging the appropriate genotypes until use. All transgenic animals were treated for seven days with doxycycline by providing doxycycline hyclate chow (Envigo, TD.120769), delivering a daily dose of 2-3 mg doxycycline based on consumption of 4-5 grams each day. On the sixth and seventh days, doxycycline hyclate (Thermo Fisher ICN19895510) was injected intraperitoneally at 50 mg/kg in 0.9% NaCl. MuSCs were isolated on the eighth day based on immunophenotype via FACS as described above.

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Brett J.O., Arjona M., Ikeda M., Quarta M., de Morrée A., Egner I.M., Perandini L.A., Ishak H.D., Goshayeshi A., Benjamin D.I., Both P., Rodríguez-Mateo C., Betley M.J., Wyss-Coray T, & Rando T.A. (2020). Exercise rejuvenates quiescent skeletal muscle stem cells in old mice through restoration of Cyclin D1. Nature metabolism, 2(4), 307-317.

Publication 2020

doxycycline hyclate chow doxycycline hyclate

0 9 nacl Animals Ccnd1 Doxycycline Doxycycline hyclate Exon Experimental animals Genotypes Heterozygous Human Immunophenotype Ires Mouse Mutation Pax7 Polyadenylation Tail Transgenic animals Wild animals

Corresponding Organization :

Other organizations : Stanford University

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Variable analysis

independent variables

Presence or absence of the K112E mutation in the human Ccnd1 transgene under TRE regulation

dependent variables

MuSCs isolated on the eighth day based on immunophenotype via FACS

control variables

All transgenic animals were treated for seven days with doxycycline by providing doxycycline hyclate chow (Envigo, TD.120769), delivering a daily dose of 2-3 mg doxycycline based on consumption of 4-5 grams each day
On the sixth and seventh days, doxycycline hyclate (Thermo Fisher ICN19895510) was injected intraperitoneally at 50 mg/kg in 0.9% NaCl

controls

Positive control: TRE-Ccnd1^WT mice transgenic for wild-type human Ccnd1 under TRE regulation
Negative control: Not explicitly mentioned

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