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 …
cells in Lgr5-eGFP-IRES-CreERT2 mice. In contrast, only 1.7 ± 0.30% of Bmi1+ ISCs, identified by the crypt Rosa-YFP+ signal after 1.5-d tamoxifen exposure in Bmi1-CreER; Rosa26-YFP mice, incorporated EdU (Fig. 1 A–F and N). To examine the relative contribution of Lgr5 vs. Bmi1 ISCs to tissue regeneration under steady-state conditions
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AX10 (Standard) Animal Health Reports Facility Barrier Level Descriptions Pricing & Availability Available Live mice available in varying quantities. 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 Animals Lgr5 Egfp Ires Creert2 Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. 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).
Contact us Gene-editing proposal Back to top Heterozygous mice harbor an Lgr5-EGFP-IRES-creERT2 "knock-in" allele that both abolishes Lgr5 gene function and expresses EGFP and CreERT2 fusion protein. When these mice mice are bred with mice containing a loxP-flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Lgr5-expressing cells of the offspring.
quence of adenoma formation, we generated Lgr5-EGFP-. IRES-creERT2; APCflox/flox mice. Intraperitoneal injection of tamoxifen induced acute loss of APC in a
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 120,429 base pairs of the human FOXF2 locus. 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.
Additionally, we exposed Lgr5-EGFP-IRES-CreERT2 mice to azoxymethane/dextrane sodium sulfate (AOM/DSS), which induces inflammation-driven colon tumors. Tumors were then flow-sorted into fractions of epithelial cells that expressed high or low levels of Lgr5 and were molecularly characterized using gene expression profiling and array comparative genomic hybridization.
The medium was removed and the Matrigel containing organoids were washed one time gently with warm PBS. Additionally, we exposed Lgr5-EGFP-IRES-CreERT2 mice to azoxymethane/dextrane sodium sulfate (AOM/DSS), which induces inflammation-driven colon tumors. Tumors were then flow-sorted into fractions of epithelial cells that expressed high or low levels of Lgr5 and were molecularly characterized using gene expression profiling and array comparative genomic hybridization. 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 Lgr5 EGFP‐Ires‐CreERT2 knock‐in mice (Barker et al., 2007) and Rosa26‐LacZ mice were obtained from the Jackson Laboratory. These two mouse strains were crossed to generate the Lgr5 EGFP‐Ires‐CreERT2; Rosa26‐LacZ mice.
Submit. Contact us Gene-editing proposal Back to top
Heterozygous mice harbor an Lgr5-EGFP-IRES-creERT2 "knock-in" allele that both abolishes Lgr5 gene function and expresses EGFP and CreERT2 fusion protein. When these mice mice are bred with mice containing a loxP-flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Lgr5-expressing cells of the offspring. 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
When using the Lgr5-EGFP-IRES-creERT2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #008875 in your Materials and Methods section.
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Lgr5 Egfp Ires Creert2, supplied by The Jackson Laboratory, used in various techniques.
The Lgr5 EGFP-Ires-CreERT2 knock-in mice (Barker et al., 2007) and Rosa26-LacZ reporter mice were obtained from The Jackson Laboratory. 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). Long-term culture of adult progenitor cells in 3D is a recently emerging technology that inhabits the space between 2D cell lines and organ slice culture. Adaptations to defined media components in the wake of advances in ES and iPS cell culture has led to the identification of conditions that maintained intestinal cell progenitors in culture.
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Methods Studies were performed in Lgr5−EGFP-IRES-creERT2 , Bmi1-CreERT2 , Hopx-CreERT2 , and TRE-H2BGFP::Hopx-CreERT2::lox-stop-lox-tdTomato
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. This revealed the presence of S-phase (i.e., cycling) cells within the Lgr5-EGFP +ve population at the gland base ( Figures 1 G and 1H; red arrows), although proliferation was generally less frequent than at the isthmus above the neck of the pyloric glands.
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We crossed Lgr5-EGFP-IRES-CreERT2 knock-in and ROSA26-tdTomato (LRT) mice, and treated them with or without 4-hydroxytamoxifen (4-OHT), to determine
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.
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Lgr5/GFP+ve cells were isolated using flow cytometry. Lgr5-eGFP-IRES-CreERT2 and Bmi1-CreER; Rosa26-YFP mice were used to compare the basal proliferation status of Lgr5 + vs. Bmi1 + ISCs during homeostasis. 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. On average, 3.72 ± 1.02 Lgr5-EGFP +ve cells were present in isolated pyloric glands (see Figure 1 E for examples). Lgr5-EGFP-Ires-CreERT2;R26R-Confetti double transgenic mice were treated with vehicle (n = 8) or G007-LK (n = 5) and tamoxifen according to the two administration schemes outlined in Fig. 1d, f.
15). All mice were crossed onto a C57BL/6 background for at least 10 generations. 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 Transcriptional differences between ISCs and their daughter cells can be explored by use of the Lgr5‐EGFP‐ires‐CreERT2 knock‐in (Lgr5‐ki) mouse (Supplementary Figure S1A; Barker et al, 2007). In this mouse model, GFP expression is driven from the Lgr5 locus, leading to highest GFP levels in Lgr5 + cells (GFP high). pressesLgr5-EGFPintheLgr5EGFP-Ires-CreERT2knock-inmice.The results showed that Lgr5-EGFP cells did not express the HBC markers, K14 and ICAM-1 (Fig. 1D,E).