Lepr cre

Manufactured by Jackson ImmunoResearch

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The Lepr-Cre is a genetic tool used in molecular biology research. It is a Cre recombinase enzyme expressed under the control of the leptin receptor (Lepr) gene promoter. The Lepr-Cre allows for targeted gene manipulation in cells that express the leptin receptor.

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10 protocols using lepr cre

Generating Leptin Receptor-Cre Mice

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Both LepRCre (Jackson Laboratory, 032457) and CAG-loxp-stop-loxp-tdTomtao (Rosa26^tdTm/+) (Jackson Laboratory, 007909) strains were purchased from The Jackson Laboratory and were maintained on a C57BL/6 background. To generate LepRCre;Rosa26^tdTm/+ mice, male LepRCre mice were crossed to female Rosa26^tdTm/+ mice. All experimental procedures were approved by the Institutional Animal Care and Use Committee at Duke University.

Guo W., Spiller K.V., Tang J., Karner C.M., Hilton M.J, & Wu C. (2021). Hypoxia Depletes Contaminating CD45+ Hematopoietic Cells from Murine Bone Marrow Stromal Cell (BMSC) Cultures: Methods for BMSC Culture Purification. Stem cell research, 53, 102317.

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Genetic Manipulation of Mouse Models

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Prx1-Cre, Lepr-Cre, DMP1-Cre, and IFT20^f/f mice were ordered from The Jackson Laboratory (Bar Harbor, MA, USA). Prx1-Cre^ERT mice were a gift from Dr. Dana Graves's lab at the School of Dental Medicine, University of Pennsylvania. Adipoq-Cre mice were a gift from Dr. Ling Qin's lab at Perelman School of Medicine, University of Pennsylvania. For the Prx1-Cre^ERT;IFT20^f/f and Prx1-Cre^ERT mice, tamoxifen was administered at D9 and D13.

Li Y., Yang S., Liu Y., Qin L, & Yang S. (2022). IFT20 governs mesenchymal stem cell fate through positively regulating TGF-β-Smad2/3-Glut1 signaling mediated glucose metabolism. Redox Biology, 54, 102373.

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Murine Hematopoietic Cell Lineages

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C57BL/6NCR (strain code 556) (CD45.2+) and B6-Ly5.1/Cr (stain code 564) (CD45.1+) were purchased from Charles River Laboratories. Rosa26mTmG, Pf4-cre, Kitl^fl/fl, Lepr-cre, and Kitl^GFP/+ mice were from The Jackson Laboratories. Cxcr4^fl/fl, Lyz2-cre, Il7ra-cre, CD4-cre, and Rosa26^tdtomato/+ mice were from internal colonies. Flk2-cre mice were a gift from Dr. E. Camilla Forsberg (University of California, Santa Cruz). Although Flk2-cre transgene is inserted into Y-chromosome, our previous work showed the hematopoietic cell composition in bone marrow and secondary lymphoid organs of Flk2-cre transgenic mice is indistinguishable from their littermate controls. All mice analyzed were 8-20 week old, were maintained under specific pathogen-free conditions at Yale Animal Resources Center.

Miao R., Chun H., Feng X., Gomes A.C., Choi J, & Pereira J.P. (2022). Competition between hematopoietic stem and progenitor cells controls hematopoietic stem cell compartment size. Nature Communications, 13, 4611.

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Genetically Engineered Mouse Models

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All mice were 8–16 weeks of age and maintained on a C57BL/6 background. Prrx1-cre (stock # 005584) recombining in bone marrow mesenchymal cells of the long bones, Lepr^cre (stock # 008320) recombining in bone marrow mesenchymal stromal cells, and Rosa26^LSL-ZsGreen (stock # 007906) mice were obtained from the Jackson Laboratory. Col1a1-cre mice (with 2.3 kb promoter), recombining in mature osteoblasts, were described previously (Liu et al., 2004 (link)). The generation of Thpo^gfp, Thpo^creER, and Thpo^fl mice was described previously (Decker et al., 2018 (link)). Thpo^gfp is a null allele of Thpo. Thpo^creER knockin mice allows tracing of cells that translate THPO protein. All mice were housed in specific pathogen-free, Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC)-approved facilities at the Columbia University Medical Center. All protocols were approved by the Institute Animal Care and Use Committee of Columbia University.

Gao L., Decker M., Chen H, & Ding L. (2021). Thrombopoietin from hepatocytes promotes hematopoietic stem cell regeneration after myeloablation. eLife, 10, e69894.

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Mouse Strains for Infectious Disease Studies

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C57BL/6 mice were purchased from the Jackson Laboratories or the National Cancer Institute. Lepr-cre, Mb1^cre/+, Nr3c1^fl/fl, Ifnar^fl/fl, and Cd21-cre mice were purchased from the Jackson Laboratories. Il7^GFP/+ mice were from our internal colony. Ltb^fl/fl (Tumanov et al, 2002 (link)) and Ltbr^fl/fl (Wang et al, 2010 (link)) mice were bred at Yale Animal Resources Center. Adult males (8–13 wk) were used for L. monocytogenes infections, and adult females (8–12 wk) were used for HSV-2 infections. Male and female adult mice (8–12 wk) were used for all other experiments. All mice were maintained under specific pathogen-free conditions at the Yale Animal Resources Center and were used according to the protocol approved by the Yale University Institutional Animal Care and Use Committee.

Lim V.Y., Feng X., Miao R., Zehentmeier S., Ewing-Crystal N., Lee M., Tumanov A.V., Oh J.E., Iwasaki A., Wang A., Choi J, & Pereira J.P. (2023). Mature B cells and mesenchymal stem cells control emergency myelopoiesis. Life Science Alliance, 6(4), e202301924.

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Tissue-Specific Kdm4b Deficient Mice

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Kdm4b^f/f mice were generated as previously described (Cheng et al., 2018 (link)). We generated tissue-specific Kdm4b deficient mice by crossing Kdm4b^f/f mice with Prx1Cre (Jackson Laboratory, Cat#005584), Prx1Cre/ERT2 (Jackson Laboratory, Cat#029211), LepRCre (Jackson Laboratory, Cat#008320) and Loxp-tdTomato mice (Jackson Laboratory, Cat#007908). All mice were housed in pathogen-free facilities under 12-hr light and 12-hr dark cycle. All protocols (#2007-057, #2007-058, #2007-059, and #2013-002) were approved by the Division of Laboratory Animal Medicine of UCLA and were in accordance with US National Institute of Health guidelines. In aging studies, we utilized both male and female mice and randomly assigned to procedure groups at a sample size of ≥ 6 mice per group. For OVX, 8 mice per group were used. 7 mice per group of 8-week-old male mice were subjected to high fat diet (Research Diets, Cat#12492). PTH (1-34) (Prospec-Tany Technogene, Cat#HOR-290) was injected subcutaneously daily at a dose of 50 μg/kg body weight. However, the animal experiments were not conducted in a completely blinded fashion. The primer sequences for genotyping were listed in Table S4.

Deng P., Yuan Q., Cheng Y., Li J., Liu Z., Liu Y., Li Y., Su T., Wang J., Salvo M.E., Wang W., Fan G., Lyons K., Yu B, & Wang C.Y. (2021). Loss of KDM4B Exacerbates Bone-Fat Imbalance and Mesenchymal Stromal Cell Exhaustion in Skeletal Aging. Cell stem cell, 28(6), 1057-1073.e7.

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Genetically Engineered Mouse Lines for Developmental Biology

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All mice were housed in specific pathogen-free, Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC)-approved facilities at the Columbia University Irving Medical Center. Food and water were provided ad libitum over 12-hour light/dark cycles. Lepr-Cre (JAX stock #032457)^{66 (link)}, Gli1-CreER (JAX stock #007913)^{66 (link)}, Gli1-LacZ (JAX stock #008211)^{27 (link)}, Rosa26-loxp-stop-loxp-tdTomato (Ai9, JAX stock #007909)^{67 (link)}, Rosa26-loxp-stop-loxp-iDTR (JAX stock #007900)^{68 (link)}, Pdgfra-H2B-GFP (JAX stock #007669)^{24 (link)}, and Sox10-CreER (JAX stock #027651)^{46 (link)} mice were obtained from the Jackson Laboratory. Col1a1(HS4,5–3.2 kb)-GFP^{25 (link)} and Pdgfra-DreER^{26 (link)} were described previously. Rosa26-rox-stop-rox-eGFP (RC∷RG) mice were generated by crossing RC∷RLTG mice (JAX stock #026931)^{69 (link)} with E2a-Cre mice (JAX stock #003724)^{69 (link)} to induce germline removal of the loxp-flanked tdTomato cassette from the Rosa26 locus. Young adult mice (both males and females of 6–12 weeks old) were used in this study. All experimental protocols were approved by Columbia University’s Institutional Animal Care and Use Committee.

Moores S.L., Sabry J.H, & Spudich J.A. (1996). Myosin dynamics in live Dictyostelium cells.. Proceedings of the National Academy of Sciences of the United States of America, 93(1), 443-446.

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Investigating Penk-Positive Neurons in Mice

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All experiments were conducted with 12–20-week-old wild-type C57BL/6J, Penk-Cre (stock no. 025112), and Lepr-Cre (stock no. 008320) mice, which were attained from the Jackson Laboratory. Penk-Cre and Lepr-Cre mice for all experiments were crossed with C57BL/6J mice for several generations before use. To visualize the Penk-positive neurons, Penk-Cre mice were crossed with Ai14 mice (stock no. 007908; tdTomato reporter line; Jackson Laboratory). Male and female mice were used for all experiments, including immunohistochemistry, FISH, anatomical tracing, food intake measurements, and behavioral experiments. Mice were group housed on a 12-h light/dark cycle with standard bedding in a temperature- and humidity-controlled room (~21 °C and 42% humidity). All behavioral experimentations were performed during the light cycle. Experimenters were blinded to the group allocation and outcome assessment. All experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Virginia Polytechnic Institute and State University.

You I.J., Bae Y., Beck A.R, & Shin S. (2023). Lateral hypothalamic proenkephalin neurons drive threat-induced overeating associated with a negative emotional state. Nature Communications, 14, 6875.

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Conditional Genetic Manipulation of Mice

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Nestin-GFP, Col2a1-cre, Osx-cre, Col2a1-creER^T2, Osx-creER^T2, Col1-creER^T2, Ocn-creER^T2, Ihh-Neo/null, Runx2-LacZ/null, Ptch1-LacZ/null, Cxcl12-GFP/null mice have been described elsewhere. Tie2-cre (JAX8863), LepR-cre (JAX8320) and Rosa26-loxP-stop-loxP-tdTomato (R26R-tomato, JAX7914) were acquired from Jackson laboratory. All procedures were conducted in compliance with the Guideline for the Care and Use of Laboratory Animals approved by Massachusetts General Hospital’s Institutional Animal Care and Use Committee (IACUC). For embryonic experiments, male mutant mice were mated to female CD1 mice and the vaginal plug was checked in the morning. Pregnant mice received 1mg tamoxifen (Sigma T5648) and progesterone (Sigma P3972) i.p. For postnatal experiments, 3 days-old mice received 0.1mg of tamoxifen i.p. tamoxifen was dissolved first in 100% ethanol then in sunflower seed oil (Sigma S5007) overnight at 60C°.

Ono N., Ono W., Mizoguchi T., Nagasawa T., Frenette P.S, & Kronenberg H.M. (2014). Vasculature-associated cells expressing nestin in developing bones encompass early cells in the osteoblast and endothelial lineage. Developmental cell, 29(3), 330-339.

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Characterization of Cre Mouse Lines

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The Col10a1Cre mouse line was a generous gift from Dr. Klaus von der Mark and has been previously described (Park et al., 2015 (link)). The Col10a1CreERT2 mouse line was a generous gift from Dr. Kathryn Cheah (University of Hong Kong). The Rosa26^fs-tdTomato, Pdgfra^H2B-GFP, LeprCre, and AdipoqCre mouse lines were obtained from Jackson Laboratories (007909, 007669, 008320, and 028020, respectively). All animal work was approved by Duke University Institutional Animal Care and Use Committees (IACUC). Both sexes of mice were utilized for experiments.

Long J.T., Leinroth A., Liao Y., Ren Y., Mirando A.J., Nguyen T., Guo W., Sharma D., Rouse D., Wu C., Cheah K.S., Karner C.M, & Hilton M.J. (2022). Hypertrophic chondrocytes serve as a reservoir for marrow-associated skeletal stem and progenitor cells, osteoblasts, and adipocytes during skeletal development. eLife, 11, e76932.

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