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Intestinal Crypt Homeostasis Results from Neutral Competition between Symmetrically Dividing Lgr5 Stem Cells Hugo J. Snippert,1 Laurens G. van der Flier,1 Toshiro Sato,1 Johan H. van Es,1 Maaike van den Born,1 Carla Kroon-Veenboer, 1Nick Barker, Allon M. Klein,2, 3Jacco van Rheenen, 1Benjamin D. Simons, and Hans Clevers ,* 1Hubrecht Institute, KNAW and University Medical Center Utrecht

usingthe knock-in allele Lgr5EGFP-Ires-CreERT2,we have shown that the cell surface receptor Lgr5 (leucine-rich repeat–containing heterotrimeric guanine nucleotide–binding protein–coupled re-ceptor 5) marksnormal tissue stem cells in stom-ach, small intestine, colon, and hair follicles (3). ApcMin and Lgr5-EGFP-ires-CreERT2 mice were obtained from The Jackson Laboratory (Bar Harbor, ME). Foxf1 and Foxf2 targeted mutants have been described elsewhere.22,23 Tg(FOXF2) was generated by pronuclear injection of a linearized BAC (RP4-668J24) spanning … 2021-01-15 Lgr5-EGFP-ires-CreERT2 mice were subjected to real-time qPCR analysis. This conﬁrmed that Lgr5, Ascl2, Tnfrsf19 and Olfm4 were highly enriched in stem cells of the small intestine (Figure S1H). Only Lgr5 and Ascl2 were enriched in the GFPhi stem cells of the colon. Expression of Bmi1, another putative N et al. Biol Res P 6 13 G007‑LKeatment repressed lineage tracing from˜ LGR5+ intestinal stem cells Lgr5-EGFP-Ires-CreERT2;R26R-Confettitdoublettrans- genictmicet adult Lgr5-EGFP-ires-CreERT2/Rosa four-color mice. Confocal.

Employing Lgr5-lacZ transgenic mice and Lgr5 in situ hybridization, we found colonic epithelial stem cells (CESC 2018-02-02 · Lgr5-eGFP-IRES-CreERT2 mice were crossed with B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J mice (Jackson Laboratories) to generate the Lgr5-eGFP-IRES-CreERT2; Rosa26-CAG-tdTomato heterozygote. To examine the contribution of Lgr5 ISC to tissue regeneration under steady-state conditions, lineage tracing was induced by tamoxifen administration in Cre reporter mice to mark the ISC and their tiation. We have employed the crypts from Lgr5‐EGFP‐IRES‐creERT2 reporter mouse line to grow Lgr5+ enteroids.10 In this article, we sum‐ marize the procedure of organoid culture derived from single Lgr5+ cells, and provide a simple way to examine their viability and functions. An overview of the experimental workflow is given in Figure 1.

b, Frequency at which the blue 2021-01-15 · Lgr5-eGFP-IRES-CreERT2 mice were crossed with B6.Cg-Gt (ROSA)26Sortm9(CAG-tdTomato)Hze/J mice (Jackson Laboratories) to generate Lgr5-eGFP-IRES-CreERT2; Rosa26-CAG-tdTomato heterozygote. Involvement Lgr5 RSCs in rectal epithelial regeneration was examined by lineage tracing assay. To this end, we first established a sorting strategy that would enable the isolation of pure populations of Lgr5 + E10.5-E12.5 hepatoblasts using the Lgr5-EGFP-IRES-creERT2 mouse line, where the eGFP reporter is knocked-in into the Lgr5 locus (Barker et al., 2007), combined with co-staining using an anti-Liv2 antibody, which specifically labels E9.5-E13.5 liver progenitors (Nierhoff et al Se hela listan på blog.csdn.net Lgr5-EGFP-IRES-creERT2基因敲入工具鼠索取资料欢迎您在此索取产品资料，联系订购产品，提交后您的需求信息将立即自动发送到供应商的E-mail邮箱，同时也将抄送一封到您的邮箱，感谢您的惠顾！ with WT Lgr5-EGFP-ires-creERT2 mice and by multiple generation selective breedings obtaining mice that were WT, CD44 /,or TLR4 / and heterozygous for the Lgr5-EGFP-ires-creERT2 re-porter.

Lgr5 Egfp Ires Creert2, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 89/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more

ZERO BIAS - scores, article reviews, protocol conditions and more 2018-01-31 · Inhibition of BMP signaling in Lgr5 cells was achieved by crossing Lgr5-EGFP-ires-CreERT2 (Lgr5-Cre) mice to mice with floxed alleles of BMP receptor 1A (Lgr5-Cre;Bmpr1aflox/flox mice). Lgr5/GFP+ve cells were isolated using flow cytometry. 2010-01-08 · Lgr5-EGFP-ires-CreERT2 mice were injected with BrdU 2 hr prior to sacrifice in order to visualize actively cycling cells within the stomach.

Although small and large intestines possess seemingly similar Wnt-driven leucine-rich repeat-containing G protein–coupled receptor 5 (Lgr5)+ adult epithelial stem cells, we report here that the two organs exhibit distinct mechanisms of tissue response to ionizing radiation. Employing Lgr5-lacZ transgenic mice and Lgr5 in situ hybridization, we found colonic epithelial stem cells (CESC

We used short-term tamoxifen exposure, for induction of Cre-mediated recombination, to selectively mark Bmi1 + ISCs in vivo. By using the knock-in allele Lgr5EGFP-Ires-CreERT2, we have shown that the cell surface receptor Lgr5 (leucine-rich repeat–containing heterotrimeric guanine nucleotide–binding protein–coupled Throughout the pyloric region of adult stomachs, both the Lgr5-LacZ (Figures 1 A and 1B) and Lgr5-EGFP-ires-CreERT2 (Figures 1 C–1E) reporters were exclusively expressed in cells occupying the very base of the glands.

Neonate (P1) Lgr5-EGFP-ires-CreERT2/R26RLacZ mice were given a single 100 μg dose of 4-hydroxytamoxifen to activate the Cre enzyme and, as a consequence, permanently switch on the lacZ reporter gene within renal Lgr5 +ve cells.
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The mammalian intestinal epithelium is endowed with a high cell turnover sustained by a few stem cells located in the bottoms of millions of crypts. Until recently, it was generally assumed that the extreme sensitivity to DNA damaging agents leading to cell death and the asymmetric mode of chromosome segregation of intestinal epithelial stem cells prevented the illicit survival of mutated stem Lgr5-EGFP-IRES-CreERT2/Rosa-LSL-tdTomato mice.

To induce Cre recombinase in Lgr5 EGFP-Ires-CreERT2 ; Rosa26-LacZ mice , tamoxifen (Sigma) was dissolved in corn oil and injected intraperitoneally from postnatal day 3 to 4 (P3–P4; 0.3 mg/g body weight). Young (2–3-month-old) and old (> 19-month-old) Lgr5-EGFP-IRES-creERT2 mice (The Jackson Laboratory) were total-body irradiated with or without 10 Gy of X-rays and sacrificed 8 h later. Swiss roll sections of paraffin-embedded intestines were prepared and immunohistochemically analyzed. May 8, 2014 GFP knockin allele: Yy1f/f;Vil-creERT2; Lgr5-EGFP-IRES-.
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H pylori colonize and manipulate the progenitor and stem cell compartments, which alters turnover kinetics and glandular hyperplasia. Bacterial ability to alter the stem cells has important implications for gastrointestinal stem cell biology and H pylori-induced gastric pathology.

Only Lgr5 and Ascl2 were enriched in the GFPhi stem cells of the colon. Expression of Bmi1, another putative N et al. Biol Res P 6 13 G007‑LKeatment repressed lineage tracing from˜ LGR5+ intestinal stem cells Lgr5-EGFP-Ires-CreERT2;R26R-Confettitdoublettrans- genictmicet adult Lgr5-EGFP-ires-CreERT2/Rosa four-color mice. Confocal. analyses of the clonal output were performed at 14 days, 2 months, 6 months, and 12 months after the ﬁnal tamoxifen.