Adult C57/BL6J mice weighing 20–25 g were purchased from the experimental animal center of Medical College at Xi’an Jiaotong University. Bmal1flox/flox mice were purchased from The Jackson Laboratory (Stock No: 007668). The animals were housed under controlled humidity (55 ± 5%) and temperature (20 ± 2 °C) conditions with a normal 12-h light/12-h dark cycle. Water and food were available ad libitum. All animal procedures were carried out in line with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee. All procedures performed in studies involving animals were approved by the Ethics Committee of Xi’an Jiaotong University (Ethics and Science # G-83) in full accordance with the ethical guidelines of the National Institutes of Health for the care and use of laboratory animals. All efforts were made to minimize suffering and the number of mcie used in the experiments.
Bmal1flox flox mice
Manufactured by Jackson ImmunoResearch
Bmal1flox/flox mice are genetically modified mice that have the Bmal1 gene flanked by loxP sites. This allows for the conditional deletion of the Bmal1 gene in a time- and tissue-specific manner using Cre recombinase technology.
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2 protocols using bmal1flox flox mice
1
Bmal1 Deletion in C57/BL6J Mice
2
Circadian Knockout Impacts Intestinal Tumorigenesis
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Mice were housed on a 12-hour light/12-hour dark cycle with ad libitum food in a barrier facility. Apcmin mice (#002020; Jackson Labs, Bar Harbor, ME) were crossed with Bmal1+/- null mutant mice (#009100; Jackson Labs). Progeny were back-crossed for 7 generations to generate a genetically pure strain before experiments began. Bmal1flox/flox mice (#007668; Jackson Labs) were crossed with VillinCre/+ mice (#021504; Jackson Labs), or Apcmin mice, then the 2 strains were crossed to obtain the Apcmin/+; Bmal1flox/flox and Apcmin/+; VilCre/+; Bmal1flox/flox for the cKO experiments. Genotyping was performed according to protocols available at Jackson Labs (www.jax.org ). All animals were tested for tumor initiation at 1 or 3 months of age. Mice maintained under constant light (LL) were housed in these conditions from 21 days of age to 3 months of age. Light levels were approximately 700 Lux, measured at the back of each cage, and all objects were removed so that light fully penetrated throughout each cage. Mice were maintained under Canadian Council for Animal Care guidelines (University of Windsor: AUPP 17-21).
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