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Gt rosa 26sortm1 cag brainbow2.1 cle j

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The Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J is a laboratory mouse strain that expresses a Cre-dependent Brainbow2.1 reporter allele at the ROSA26 locus. The Brainbow2.1 reporter allows for the stochastic expression of different fluorescent proteins in individual cells, enabling visualization of cellular morphology and connectivity.

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5 protocols using gt rosa 26sortm1 cag brainbow2.1 cle j

1

Inducible Pax2-Driven Lineage Tracing

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Pax2.rtTA;TetO.Cre;mT/mG mice or Pax2.rtTA;TetO.Cre;R26.Confetti mice were developed by crossing the mT/mG reporter strain B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J or the Confetti strain Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J with the TetO.Cre strain B6.Cg-Tg(TetO-Cre)1Jaw/J, both purchased from Jackson Laboratory. Double-transgenic mice were then crossed with Pax2.rtTA mice (Burger et al., 2011 (link)) to obtain a triple-transgenic inducible mouse model with a 100% C57Bl/6 background (Figures S1A and S2A). For information on genotyping, induction procedures, and tissue processing, see Supplemental Experimental Procedures.
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2

Clonal Analysis of c-Kit+ Olfactory Lineage

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All experimental procedures were approved by the University of Miami Institutional Animal Care and Use Committee, and were performed in full compliance with the NIH Guidelines for the Care and Use of Laboratory Animals. The c-KitCreERT2/+ mouse line was provided by Dr. Dieter Saur (Klein et al., 2013 (link)). The multicolor Cre reporter, Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J (Stock Number: 013731), abbreviated here as R26R-Confetti, was obtained from the Jackson Laboratory (Bar Harbor, ME). For conditional labeling of c-Kit+-derived cells, c-KitCreERT2/+ mice were crossed with R26R-Confetti mice. PCR genotyping for c-KitCreERT2/+ was performed as described (Klein et al., 2013 (link)); R26R-Confetti mice were bred as homozygotes. In initial experiments, tamoxifen (Sigma, St. Louis, MO) 20 mg/mL in peanut oil (Sigma) was given at 2 mg intraperitoneally at designated times to c-KitCreERT2/+; R26R-Confetti adults, or 0.2 mg to postnatal mice. For clonal analysis of the c-Kit+ olfactory lineage, animals were given a single low dose of tamoxifen, determined empirically to yield sufficiently sparse labeling: 1 mg for adult mice, and 0.0125 mg for neonates.
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3

Transgenic Mice for Lineage Tracing

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The Tg(Pax8-rtTA2S*M2)1Koes/J (Pax8-rtTA) Tg(tetO-Cre)1Jaw/J (Tre-Cre) mice (Perets et al., 2013 (link)), Foxj1tm1.1(cre/ERT2/GFP)Htg/J (FoxJ1CreERT2::GFP; JAX stock no. 027012),Gt(ROSA)26Sortm9(CAG-tdTomato)Hze (Ai9) mice (JAX stock no. 007909) and Confetti mice [Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J; JAX stock no. 013731] were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). The Trp53loxP/loxP and Rb1loxP/loxP mice, which have Trp53 and Rb1 genes, respectively, flanked by loxP alleles, were a gift from Dr Anton Berns (The Netherlands Cancer Institute, Amsterdam, The Netherlands). All the experiments and maintenance of the mice followed the recommendations of the Guide for the Institutional Laboratory Animal Use and Care Committee.
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4

Transgenic Mouse Strains for Immunology

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C57BL/6J (B6), B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J (mTomato), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I), C57BL/6-Prf1tm1Sdz/J (PRF−/−), and Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J (Confetti) mice were purchased from the Jackson Laboratory. Blimp1-YFP (provided by M. Nussenzweig, Rockefeller University), actin-mCerulean, actin-mOrange, actin-YFP, IFNAR−/− (provided by J. Sprent, formerly at the Scripps Research Institute), KL25 H (provided by H. Hengartner, University of Zurich), KL25 H+L (provided by M. Iannacone, San Raffaele Scientific Institute), VI10YEN, P14, Thy1.1+ P14, SMARTA, mCerulean+ SMARTA, and mOrange+ SMARTA (all on a pure B6 background) were bred and maintained under specific pathogen–free conditions at the National Institutes of Health (NIH). The KL25 H+L mice were originally generated by P. Greenberg at the University of Washington. All mice in this study were handled in accordance with the guidelines set forth by the NIH Animal Care and Use Committee.
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5

Murine Intestinal Epithelial Lineage Tracing

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C57BL/6 mice were bred in Tierforschungszentrum (TFZ), Ulm University. Tg(Vil-cre/ERT2)23Syr, Gt(ROSA)26Sortm1(CAG-Brainbow2.1)Cle/J, and B6.129P2-Lgr5tm1(cre/ERT2)Cle/J were purchased from the Jackson Laboratory. Tg (Vil1-cre/ERT2) mice were crossed with Confetti; R26 to get vil1-cre; R26 Confetti mice and Lgr5EGFP-ires-creERT2 were crossed with Confetti; R26 to get Lgr5e-creERT2; R26 Confetti. All transgenic mice were backcrossed to C57BL/6 mice. To initiate the confetti fluorophores either in the whole intestinal epithelial cells or specifically in the Lgr5 cells, vil1-cre; R26 Confetti or Lgr5e-creERT2; R26 Confetti mice, respectively, were injected intraperitoneal with a single dose (5 mg) of tamoxifen (Sigma-Aldrich). All mice were housed in the animal barrier facility under pathogen-free conditions at the Ulm University. All mouse experiments were performed in compliance with the German Law for Welfare of Laboratory Animals and were approved by the Institutional Review Board of the Ulm University and by the Regierungspraesidium Tuebingen (state government of Baden-Württemberg), protocol number: 35/9185.81-3/1407. Housing conditions: a temperature range of 22°C ± 1°C, a relative humidity of 55% ± 10%, an air change rate of 15 times, and a light/dark change of 12/12 h. Nutrition: ssniff M-Z autoclavable complete for mice breeding (#V1124-3).
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