CD11c-cre Il27p28fl/fl mice (Lee et al., 2015 (link)) and Foxp3creIl27rafl/fl mice (Do et al., 2017 (link)) were described previously. In addition, CD4-cre (Lee et al., 2001 (link)), Lysozymecre (Clausen et al., 1999 (link)), Vil-cre mice (Madison et al., 2002 (link)), and CD45.1+ B6 mice were purchased from the Jackson Laboratory. All mice were bred and housed under specific pathogen–free conditions. 8–12-wk-old mice of both sexes were used as young mice, 6–7-mo-old mice were used as aged mice, and only WT littermates of the same gender served as controls in each experiment. All mice were maintained and handled in accordance with the Institutional Animal Care and Use Guidelines of University of California, San Diego and National Institutes of Health Guidelines for the Care and Use of Laboratory Animals and the Animal Research: Reporting In Vivo Experiments guidelines.
Vil cre mice
Manufactured by Jackson ImmunoResearch
Vil-cre mice are a type of genetically engineered mouse model that express the Cre recombinase enzyme under the control of the villin promoter. The villin promoter drives Cre expression specifically in the intestinal epithelial cells of the mouse. This allows for targeted gene manipulation and analysis within the intestinal tissue of these mice.
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3 protocols using vil cre mice
1
Genetically Modified Mouse Models
2
Intestine-specific RhoA Inactivation
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All animal experiments were carried out under protocols approved by Vall d’Hebron University Hospital Research Institute Animal Experimentation Ethics Committee. C57BL/6JJcl-CAT-RhoA-DN/4–18 mice carrying the cytomegalovirus promoter fused to a CAT gene cassette flanked by loxP sites and the dominant negative T19N mutant of RhoA were obtained from the RIKEN BioResource Research Center (Starin RBRC01295; hereafter called RhoAT19N mice).28 (link) The C57BL/6J.SJL-Tg(Vil1-cre)997Gum/J mice hemizygous for the Villin-Cre transgene expressing Cre recombinase under the control of the Villin 1 promoter (hereafter called Vil-Cre+ mice) were obtained from the Jackson Laboratory (Strain #:004586).29 (link) Hemizygous RhoAT19N mice were crossed with hemizygous Vil-Cre+ mice, resulting in intestine-specific expression of RhoAT19N in Vil-Cre+;RhoAT19N mice. Vil-Cre–;RhoAT19N mice were used as control in all the experiments. Mouse genotyping was performed with DNA prepared from the mouse ear. Specific PCR primers for RhoAT19N and Vil-Cre were used (Table S1 ). All experiments included animals from both sexes.
3
Generation of Mettl3 and Alkbh5 Knockout Mice
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Mettl3 conditional knockout mice were generated by inserting two loxp sites into the intron after the first exon and the intron before the last exon of Mettl3 using CRISPR/cas9 based genome-editing system as previously described [5] . Alkbh5 conditional knockout mice were generated by inserting two loxp sites into the introns flanking the first exon of Alkbh5 using CRISPR/cas9 based genome-editing system as previously described [36] . Genotyping of Mettl3 f/f mice, Alkbh5 f/f mice, Vil-cre mice (The Jackson Laboratory, Stock No: 021504), and Irf7 -/-mice (RIKEN BRC, RBRC01420) were confirmed by PCR using primers as list below:
The sex-, age-and background-matched littermates of the knockout or conditional knockout mice were used as the controls in the present study. All mice were on the C57BL/6 background. Mice were maintained in SPF conditions under a strict 12 h light cycle (lights on at 08:00 and off at 20:00). All animal studies were performed according to approved protocols by the Ethics Committee at the University of Science and Technology of China (USTCACUC202101016).
The sex-, age-and background-matched littermates of the knockout or conditional knockout mice were used as the controls in the present study. All mice were on the C57BL/6 background. Mice were maintained in SPF conditions under a strict 12 h light cycle (lights on at 08:00 and off at 20:00). All animal studies were performed according to approved protocols by the Ethics Committee at the University of Science and Technology of China (USTCACUC202101016).
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