Ert2 estrogen. pA, a polyA sequence from SV40 virus.

Ert2 estrogen The estrogen receptor ligand activated Cre-ERT2 to express EML4-ALK gene in the lungs. In the split-CreERT2 system, Cre-mediated DNA recombination is controlled by two expression cassettes as well as the time of tamoxifen Aug 20, 2022 · Much is known about estrogen action in experimental animal models and in human physiology. Induction in adult mice avoids genetic alterations early on in development, which could lead to unwanted, potentially harmful effects. Dec 17, 2024 · Mutation details: The transgene consists of a tamoxifen-inducible cre recombinase driven by the 2. cific Cre and Cre/ERT2 alleles have some degree of unreported activity, inconsistent recombination efficiency, and parent of Selective estrogen receptor modulators (SERMs) are an increasingly important therapeutic modality that are used by clinicians on a daily basis. Mar 21, 2009 · recombinase fused estrogen receptor with mutated ligand binding domain is bound to the heat shock protein (Hsp90) and inhibited from the entering the nucleus. We have generated a broadly expressed inducible ERT2-GAL4 zebrafish line using the ubiquitin (ubi) enhancer. Estrogen/GPR30 or estrogen/ERα complex mediated modulation of Src/EGFR interaction leading to To explore the function of genes expressed in adult mouse nociceptive neurons, we generated heterozygous knock-in mice expressing the tamoxifen-inducible Cre recombinase construct CreERT2 downstream of the Na V 1. 129S-Tg (UBC-cre/ERT2)1Ejb/J (hereafter abbreviated as Cre – ERT2), a fusion of a mutated estrogen receptor and Cre recombinase, was Apr 12, 2018 · Cre ERT2 is a recombinant fusion protein of the Cre DNA recombinase and a mutant ligand-binding domain of the human estrogen receptor. , Wagner, J. Jun 1, 2004 · Purpose: Approximately two-thirds of breast cancer patients respond to endocrine therapy, and this population of patients is estrogen receptor (ER) positive. Nov 14, 2023 · 1292 volume 10 number 12 december 2009 nature immunology Articles Most successful vaccines are antibody based, and some memory B cell clones can persist throughout life along with specific circulating Jan 20, 2025 · ERT2: estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A) 6: ERT2: En2: human estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A, phage P1 P1 codon improved Cre (iCre), mouse PGK promoter, Streptomyces alboniger puromycin resistant gene, Download scientific diagram | Generation of Cre–ERT2 mT/mG mice and inducible Ece1 Knockout (KO) mice. One of the main advantages of the system is that Cre-mediated recombination can be controlled in time through Tamoxifen To explore the function of genes expressed in adult mouse nociceptive neurons, we generated heterozygous knock-in mice expressing the tamoxifen-inducible Cre recombinase construct CreERT2 downstream of the Na(V)1. 5 The molecular chaperone Hsp90 interacts with unliganded steroid hormone receptors and regulates their activity. The resulting HA-ERT2-I-Ppo-I cDNA was inserted into the STOP-eGFP-Rosa26 targeting vector (Addgene plasmid 11739, ) upstream of the IRES-eGFP cassette. The gene product contains an N-terminal DNA binding domain and C-terminal ligand binding domain and is localized to the nucleus, cytoplasm, and mitochondria. Dec 19, 2015 · An improved estrogen receptor nuclear translocation domain (ERT2) with N-terminal HA tag was cloned upstream of the I-PpoI cDNA (kind gift from M. Jan 17, 2022 · Based on the dimorphic difference in body size at 3 months in Acan-Cre ERT2;Stat3 fl/fl females and males, we hypothesized that estrogen may regulate Stat3 levels and/or activity. As an example, mice Dec 5, 2022 · The first ERα PROTAC, developed by Crews, Deshaies and co-workers [17] was based on the natural hormone, namely 17β-estradiol (E2) and a phosphopeptide to recruit the E3 ligase. & Chambon, P. receptor and Cre recombinase, was engineered to be more. However, a significant proportion of patients do not respond to hormone therapy. May 11, 2017 · A variety of conditional knock-out mice relying on Tamoxifen-driven ERT2/Cre -mediated recombination are available and have been used to (SBE). We show Nov 26, 2015 · By attaching Cre to a mutated estrogen receptor (ERT2, which binds TAM but not endogenous estrogens; Cre-ERT2), it is possible to control precisely when a DNA sequence can be excised. Unlike estrogen antagonists in human breast cancer, it is an estrogen agonist in mouse bone [9]. 1 a). The second generation of inducible Cre-ERT recombinase, hemizygous B6. Allele Symbol Allele Name Common Names Promoter Diseases Related to This Gene; ERT2: estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A) 5: ERT2: gastrointestinal stromal tumor()idiopathic hypereosinophilic syndrome()polyps, multiple and recurrent inflammatory fibroid, gastrointestinal() Sep 15, 2020 · Genetically engineered mouse models through gene deletion are useful tools for analyzing gene function. 104 Immunostaining for estrogen receptor (ESR) showed at E9. The effects of E2 on cells are mediated by the transcription factors and estrogen receptor (ER)α and ERβ that are encoded by distinct genes. The discovery of the Von Hippel–Lindau tumour suppressor (pVHL) and its activity as a degrader Download scientific diagram | Localisation and activity of GAL4-VP16/estrogen receptor LBD fusion proteins are 4-OHT dependent. Sep 15, 2020 · Here, we generated transgenic mouse with cardiac-specific expression of Cre recombinase fused to a mutant estrogen ligand-binding domain (ERT2) on both N-terminal and C-terminal under the regulatory region of human vasoactive intestinal peptide receptor 2 (VIPR2) intron and Hsp68 promoter (VIPR2-ERT2CreERT2). Its specific role in mouse epiblast stem cell self-renewal, however, remains poorly understood. To contribute to the reduction of animals in research, Cre-negative mice from HSACreERT2 and Pax7CreERT2(FAN) breedings were used in C57Bl/6 wildtype (WT) experiments. Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox system. For tamoxifen-dependent Cre recombinase, also known as CreER recombinase, tamoxifen (TAM) is used to activate the Cre to generate time- and tissue-specific mouse mutants. Mice Thesecontradicting observations ofthechemicalpropertiesoftrans-4-OHT havedictated currentlyused laboratoryprotocolsoftrans-4-OHT handling. The transgene insert contains a fusion product involving cre recombinase and a mutant form of the mouse estrogen receptor ligand binding domain. Conditional DNA excision between two LoxP sites can be achieved in the mouse using Cre-ER T, a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER ligands. Mutation details: The mouse promoter drives cardiac-specific expression of a tamoxifen-inducible humanized cre (icre) fused to an estrogen receptor mutant (ERT2) followed by a bovine growth hormone polyadenylation sequence. Oct 22, 2019 · The CreERT2/loxP system is widely used to induce conditional gene deletion in mice. The two recombineering steps are outlined. Cg- Ndor1 Tg(UBC-cre/ERT2)1Ejb /1J When these UBC-Cre-ERT2 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 widespread cells/tissues. 2013 Jul 19;288(29):21105-21116. Estrogen is critical for skeletal homeostasis and regulates bone remodeling, in part, by modulating the expression of receptor activator of NF-κB ligand there is evidence from lineage tracing experiments employing Dmp1-Cre-ERT2 mice that the Dmp1 promoter is active in osteoblasts and their descendants 59, 60, corroborating our data. We report here the construction and characterization of a series of chimeric recombinases, each consisting of Cre fused to a mutated human oestrogen receptor (ER) ligand-binding domain (LBD). M113. The ERT2 sequence is an estrogen receptor targeting motif, whose nuclear translocation and activity are controlled by an estrogen HSPC-P, human surfactant protein C promoter; Cre-ERT2, Cre coding sequence fused with a tamoxifen-inducible estrogen receptor. [13] It is also being studied for other types of cancer. doi: 10. a A schematic showing the generation of Cre–ERT2 mT/mG mice. then I add tamoxifen to check the color change. An estrogen receptor variant (ERT2) fused to the GAL4 transcriptional activator rapidly For a more thorough introduction, check out Addgene’s Cre-lox blog post. Mar 16, 2018 · Here, we designed and vigorously optimized a series of Hybrid drug Inducible CRISPR/Cas9 Technologies (HIT) for transcriptional activation by grafting a mutated human estrogen receptor (ERT2) to multiple CRISPR/Cas9 systems, which renders them 4-hydroxytamoxifen (4-OHT) inducible for the access of genome. May 15, 2013 · We have generated an inducible system to control the timing of transgene expression in zebrafish and chick. Cases can manifest spontaneously, be inherited genetically, or be acquired We report the generation of five mouse strains with the tamoxifen-inducible Cre (Cre-ER(T) (2) ; CE) gene cassette knocked into the endogenous loci of Pax3, Myod1, Myog, Myf6, and Myl1, collectively as a resource for the skeletal muscle research community. An estrogen receptor variant (ERT2) fused to the GAL4 transcriptional activator rapidly and robustly activates transcription within 3 hours of treatment with the drug 4-hydroxy-tamoxifen (4-OHT Jun 8, 2024 · Irene Garcia-Gonzalez, Susana F Rocha, Anahita Hamidi, Lourdes Garcia-Ortega, Alvaro Regano, Maria S Sanchez-Muñoz, Mariya Lytvyn, Aroa Garcia-Cabero, Sergi Roig-Soucase, Hans Schoofs, Marco Castro, Helena Sabata, Michael Potente, Mariona Graupera, Taija Makinen, Rui Benedito, iSuRe-HadCre is an essential tool for effective conditional Nov 20, 2024 · ERT2: estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A) UN: ERT2: cre: Phage P1 Cre recombinase: UN: cre: rat Nestin promoter, Nestin 2nd intron enhancer: Phenotype; Annotation by Mammalian phenotyhpe ontology: no abnormal phenotype detected; Detailed phenotype data: Ordering Information; The inducible analog of Cre is ERT2-Cre-ERT2. Although several Cre-loxP-based gene knockout mouse models have been generated for the study of gene function in alveolar epithelia in the lung, their applications are still limited. 1(cre/ERT2)Bzsh allele was generated by homologous recombination to replace exon 1 of the Nfatc1 locus with Cre/ERT2 cDNA. , 2012; Zhu et al. 5, 1, 2, 4, and 8 h, To rapidly induce RHOX10, we chose to employ the estrogen receptor T2 (ERT2) system, which drives swift translocation of the TF of interest to the nucleus in response to 4-hydroxytamoxifen This gene encodes a member of the family of estrogen receptors and superfamily of nuclear receptor transcription factors. CreERT2 encodes a Cre recombinase (Cre) fused to a mutant estrogen ligand-binding domain (ERT2) that requires the presence of Nov 1, 1999 · Abstract. 5 Jul 15, 2001 · The estrogen receptor (ER) is a ligand-activated enhancer protein that is a member of the steroid/nuclear receptor superfamily. As a consequence, the recom-binase activity of Cre-ERT2 strictly depends on the addition of these chemicals to the culture One of the SELECTIVE ESTROGEN RECEPTOR MODULATORS with which came from transgenic mice with cre-ert2 recombinase. Please refer to our privacy policy for more information. Similarly to the endogenous estrogen receptor, Cre ERT2 is Sep 12, 2016 · A modified version of Cas9 with a fusion of the hormone-binding domain of the estrogen receptor allows reversible control of Cas9 activity with high efficiency at multiple loci with 4 Aug 2, 2019 · Cre에 ERT2가 붙어있는데 이게 Estrogen receptor입니다. In this transgenic model, the 4hydroxytamoxifen (4 Here, we generated transgenic mouse with cardiac-specific expression of Cre recombinase fused to a mutant estrogen ligand-binding domain (ERT2) on both N-terminal and C-terminal under the regulatory region of human vasoactive intestinal peptide receptor 2 (VIPR2) intron and Hsp68 promoter (VIPR2-ERT2CreERT2). In Goodwin et al. 1074/jbc. Using the yeast system, we sh 2 days ago · Selective estrogen receptor modulators (SERMs), also known as estrogen receptor agonists/antagonists (ERAAs), [1] [2] are a class of drugs that act on estrogen receptors (ERs). Transgene expression in endothelial cells of the liver and kidney was confirmed by immunohistochemical analysis. The viral 2A oligopeptide sequence mediates ribosomal skipping. g. 1 day ago · Tamoxifen, sold under the brand name Nolvadex among others, is a selective estrogen receptor modulator used to prevent breast cancer in women and men. Here we show that Sep 27, 2018 · The Nfatc1 tm1. 8 promoter. 2019 Genome Res. 129S-Tg(UBC-cre/ERT2)1Ejb/J Sep 17, 2020 · The inducible Cre/loxP estrogen receptor (ER) transgenic system is used to study both ubiquitous and tissue-specific gene function through the expression of Cre recombinase Mar 21, 2009 · Cre 발현 유도 이식유전자는 Cre 재조합 효 소와 돌연변이가 있는 리간드 결합 도메인을 가진 에스트로 겐 수용기(estrogen receptor, ER)가 결합된 CreER 폴리펩 타이드를 Dec 15, 2023 · The development of the Cre- loxP system, 1,2 with subsequent integration 3 and refinement 4 of a Cre bound to a mutated estrogen receptor (ERT2) allows for temporally 3 days ago · There are two different forms of the estrogen receptor, usually referred to as α and β, each encoded by a separate gene (ESR1 and ESR2, respectively). 463455. We characterized these CE strains using the Sep 25, 2012 · A) Schematic map of SPC-Cre-ERT2 expression cassette. ERα independent binding of estrogen to G-protein coupled receptor 30. , osteoporosis). The Cre-ERT2 is active only in the presence of tamoxifen. HSPC-P, human surfactant protein C promoter; Cre-ERT2, Cre coding sequence fused with a tamoxifen-inducible estrogen receptor. 7% type II alveolar epithelial cells of SPC-Cre-ERT2/ROSA26R mouse HSPC-P, human surfactant protein C promoter; Cre-ER T2, Cre coding sequence fused with a tamoxifen-inducible estrogen receptor. pA, a polyA sequence from SV40 Jul 19, 2013 · Estrogen receptor α L543A,L544A mutation changes antagonists to agonists, correlating with the ligand binding domain dimerization associated with DNA binding activity J Biol Chem. A frt-flanked neomycin Jul 10, 2001 · The estrogen-related receptors (ERRα, ERRβ, and ERRγ) form a family of orphan nuclear receptors that share significant amino acid identity with the estrogen receptors, but May 14, 2014 · The effect of PTEN loss in the SEZ/SVZ progenitors has also been explored using three estrogen receptor T2 (tamoxifen-inducible) Cre (ERT2-Cre) lines controlled by the Nestin promoter (Nestin-ERT2) (Bonaguidi et al. We previously reported that, in male mice, cold-induced beige adipocyte formation begins to decline around 6 months of age and Flow chart of creating a Cre-ER T2 knock-in targeting vector. Two genes encode mammalian ER: ERalpha and ERbeta. The recombinase is activated by 49OHT but not by endogenous estrogens because of specific point mutations in the LBD (11, 12). CreERT2 has high affinity for the 4-hydroxytamoxifen (4-OHT) but not for the endogenous Nov 1, 2017 · Cre-ERT2 is a fusion protein between the recombinase Cre and the ligand-binding domain of the estrogen receptor, which has been mu-tated to bind only artificial estrogens, such as tamoxifen and hydroxytamoxifen (OHT) [12]. Estrogen receptor-α (ER) drives tumor development in ∼75% of breast cancer cases, and treatments directly targeting ER (e. coli Neomycin resistance gene, bovine polyA signal, yeast FRT (flipase recombination target) sites: Research application: Apr 15, 2002 · In recent years, the Cre integrase from bacteriophage P1 has become an essential tool for conditional gene activation and/or inactivation in mouse. CreERT2 is Dec 17, 2024 · Mutation details: The Nestin-cre/ERT2 transgene was designed with the rat nestin (Nes) promoter driving expression of a CreERT2 fusion gene (cre/ERT2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand-binding domain). This article reviews the mechanisms of estrogen activity in animals and humans and the role of its two receptors α and β in terms of structure and mechanisms of action in various tissues in health and in relationship with human pathologies (e. ER hormone responsiveness is widely believed to be influenced by enhanced cross-talk of ER with overexpressed human epidermal Oct 29, 2020 · Oestrogen receptor-α (ERα) shuttles continuously between the nucleus and the cytoplasm, and functions as an oestrogen-dependent transcription factor in the nucleus and as an active mediator of Feb 2, 2021 · Introduction. Based on these Cre-lox recombination principles, scientists have developed constructs to activate/inactivate genes when Cre is present. Mutations in the estrogen receptor have long been Dec 16, 2023 · The development of the Cre-loxP system,1,2 with subsequent integration3 and refinement4 of a Cre bound to a mutated estrogen receptor (ERT2) allows for temporally controlled induction of cre recombination with tamoxifen (Tam) administration. Jul 20, 2021 · Both Rhox10-ERT2 and Control-ERT2 transduced cells were incubated with 4OH-TAM for 0, 0. Key genes involved in proliferation and tumor progression are transcriptionally regulated by ERα making it an important Feb 23, 2021 · Estrogen receptor-α (ER) drives tumor development in ∼75% of breast cancer cases, and treatments directly targeting ER (e. An estrogen receptor variant (ERT2) fused to the GAL4 transcriptional activator rapidly and robustly activates transcription within 3 hours of treatment with the drug 4-hydroxy-tamoxifen (4-OHT) in tissue culture and transgenic zebrafish. Activation of ESR1 by 17β-estradiol (E2) and ESR1-selective agonist PPT increased CCND1 expression, and this effect was dependent on NF-kB activation. In this study, we developed a SPC-Cre-ER(T2) mouse model, in which a tamoxifen-inducible Cre recombinase (Cre-ER(T2)) Dec 20, 2024 · Gene; Gene Symbol Gene Name Chr. The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor; which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT) and, with lesser sensitivity, ICI 182780. The iCreER fusion gene used here has an improved/optimized variant of Cre recombinase (iCre) that is Abstract. @article Endoxifen is established as potent and reproducible complementary compound to 4-OHT to control ERT2 domain fusion proteins in vivo, and provide a framework for efficient chemically controlled recombination Oct 25, 2009 · Cre-ERT2 replaces Aicda exon 2 and is fused with sequence Feil, R. TAM is a potent CreER system inducer; however, TAM is also an active selective estrogen receptor modulator (SERM) that can influence bone homeostasis. Western Blot Oct 1, 2009 · We generated a single adenoviral vector containing a tet-inducible, composite SacI restriction endonuclease/estrogen receptor (ERT2) gene, and a constitutively expressed reverse transactivator The non-overlapping functions of the two estrogen receptor subtypes, ERα (Estrogen Receptor α)and ERβ (Estrogen Receptor β), in tumor cells have been studied extensively. Most mutations localise to just three residues at, or near, the The transgene expresses an optimized, Flp-estrogen receptor fusion protein (Flpo-ERT2) under the control of a loxP-stop-loxP cassette. 5, 1, 2, 4, and 8 h, respectively. The therapeutic efficacy of tamoxifen (TAM) in cancer therapy is thought to arise primarily from its ability to compete with estrogens for binding to the estrogen receptor (ER). The. In the Cre/loxp system, TAM binds to the ligand pocket of ER-α in another conformation as an inducer to activate Cre recombinase at specific times and inactivate the target gene [10]. Aug 28, 2020 · The Cre-ERT2 system uses the estrogen receptor ligand-binding domain to allow induction of Cre activity in response to tamoxifen, allowing for temporal control of recombination (9, 17). Sep 1, 1997 · For the present study, we generated a new transgenic mouse model with inducible SIRT6 overexpression in the RPE. the result is quite These tools allow the temporal control of autophagy via the administration of tamoxifen and spatial control via tissue or cell-specific ERT2-Gal4 driver lines and will enable the investigation of how cell- or tissue-specific changes in autophagic flux affect processes such as aging, inflammation and neurodegeneration in vivo. , aromatase inhibitors, tamoxifen, To achieve spatial-temporal control, the Cre recombinase gene is fused with a human estrogen receptor (ER) domain and expressed under tissue-specific promoter, resulting in 2 days ago · Cre recombinase consists of 343 amino acids that form two distinct domains. The creERT2 fusion gene (cre/ERT2) is cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand-binding domain. In this system, genes modified by loxP sites are altered first upon expression of Cre. CreERT2 encodes a Cre recombinase (Cre) fused to a mutant estrogen ligand-binding domain (ERT2) that requires the presence of Aug 29, 2013 · Wnt/β-catenin signalling has a variety of roles in regulating stem cell fates. We have recently characterized a new Feb 21, 2019 · However, with age, a muscle phenotype was apparent in the Sost-ERT2 Cre mice a tamoxifen-inducible Cre was generated in which a mutated estrogen ligand-binding domain was used. ER operates as part of a transcriptional complex, coordinating with several other proteins to access chromatin and in turn regulate gene A Rosa-CAG targeting vector was designed to insert cassette containing (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), loxP-site, Dre recombinase fused with a mutant form of estrogen receptor hormon-binding domaine C (ERT2), STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), second Apr 25, 2024 · The cerebrovascular endothelial cells with distinct characteristics line cerebrovascular blood vessels and are the fundamental structure of the blood–brain barrier, which is important for the development and homeostatic May 18, 2012 · The Cre recombinase fused to a mutant ligand-binding domain of the estrogen receptor (CreERT2) circumvents this problem , . Previous studies have suggested that hsp90 interacts with large portions of the estrogen receptor (ER) ligand-binding domain and sequences of the receptor required for stable DNA b Jan 10, 2018 · Next, we fused a variant of estrogen receptor (ERT2) 27 or glucocorticoid receptor (GR) with Cas9 to investigate ligand dependent nuclear shuttling of Cas9 upon Dec 5, 2016 · 17β-Estradiol (E2), as the main circulating estrogen hormone, regulates many tissue and organ functions in physiology. Jul 1, 2024 · The estrogen-dependent role of TNFR1-mediated supraspinal neuronal circuitry in CNP remains unknown. pA, a polyA sequence from SV40 virus. The pound symbol (#) is used when no line is specified and/or lines are pooled. For example, the activity of many proteins fused to the hormone-binding domain of the Jul 7, 2021 · Both Rhox10-ERT2 and Control-ERT2 transduced cells were incubated with 4OH-TAM for 0, 0. , 5. Upon administration and binding of estrogen antagonist, tamoxifen (TM) or 4-hydroxytamoxifen (4-OHT) to the mutant estrogen receptor, (B) Mice expressing LoxP-flanked mTOM stop cassette and a downstream mEGFP transgene under R26 locus were bred with mice expressing R26-regulated Cre recombinase gene fused with the gene encoding ligand-binding domain (LBD) of human estrogen receptor T2 (Cre-ERT2) to generate F1 progeny expressing single copies of Cre-ERT2 and mTOM/mEGFP transgene The transgene expresses an optimized, Flp-estrogen receptor fusion protein (Flpo-ERT2) under the control of a loxP-stop-loxP cassette. This inducible system has cre fused to a mutated estrogen receptor (ERT2) such that Cre/ERT2 is normally sequestered in the cytoplasm and inactive, but, when tamoxifen binds ERT2, the cre fusion protein translocates to the nucleus, where it is active (Indra et al. Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding Oct 15, 2020 · Myh11-Dre ERT2 mice were made by Cyagen Laboratory using traditional transgenic mice generation methodologies as previously described. Unfortunately, clinicians have a limited understanding regarding the underlying mechanism(s) of how SERMs function and their increasingly useful role in the treatment of estrogen-responsive target tissues such as the Aug 28, 1997 · Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains. A number of tissue-specific Cre-ERT2 mouse models have been reported [7,8,9]. The transgenic mice were given tamoxifen at days 3 and 6 by intraperitoneal injection. (A-I) Forced expression of ERT2-GAL4 (schematised, top) in HEK293 Feb 14, 2019 · This inducible system has cre fused to a mutated estrogen receptor (ERT2) such that Cre/ERT2 is normally sequestered in the cytoplasm and inactive, but, when tamoxifen binds ERT2, the cre fusion protein translocates to the nucleus, where it is active (Indra et al. Duration of tamoxifen treatment influences efficiency of recombination, which needs to be considered when interpreting data obtained By attaching Cre to a mutated estrogen receptor (ERT2, which binds TAM but not endogenous estrogens; Cre-ERT2), it is possible to control precisely when a DNA sequence can be excised. The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17beta-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Specificity and efficiency of Cre activity was documented by crossing VECad-Cre-ERT2 with the ROSA26R reporter The estrogen receptor (ER) is a transcription factor that binds to a specific DNA sequence found in the regulatory regions of estrogen-responsive genes, called the estrogen response . Authors Yukitomo Dec 20, 2024 · Gene; Gene Symbol Gene Name Chr. Vectors, cassettes, and pertinent elements of the constructs are as depicted and labeled; oligos are short lines; squares, restriction enzyme site overhangs; p, phosphorylated ends. Mar 14, 2018 · Cre ERT2 is silent but activates upon estrogen analog tamoxifen administration. The transgene consists of the human vWF promoter with the microvascular selective 734 base pair sequence to drive Cre recombinase fused to a mutant estrogen ligand-binding Sep 25, 2012 · We found that Cre-ERT2 was expressed in 30. Localized at the peri-membrane, mitochon Dec 17, 2024 · Mutation details: The transgenic construct encodes a tamoxifen-inducible fusion protein consisting of cre recombinase and a mutated human estrogen receptor ligand-binding domain fused to the mouse Tek promoter and 10. efficient and specific than the original Cre–ERT. We have generated an inducible system to control the timing of transgene expression in zebrafish and chick. Dec 12, 2014 · Endocrine therapy is the mainstay of treatment in estrogen receptor-positive breast cancers and significantly reduces disease recurrence and breast cancer-related mortality. The transgene was microinjected into fertilized C57BL/6J oocytes, and line K was established. The second generation of inducible Cre-ERT recombinase, hemizygous B6. , 2011; Amiri et al. , Metzger, D. Kastan). In an earlier report, we described a fusion protein between Cre and a mutated form of the ligand binding domain of the estrogen receptor (Cre-ER TM) that renders Cre activity tamoxifen (TM) inducible, allowing for Jun 10, 2024 · Estrogen counteracts age-related decline in beige adipogenesis. Reproducible experimentation mandatesthegeneration ofone-time working aliquotsoftrans-4-OHTin DMSOorEthanol Dec 17, 2024 · Mutation details: A targeting vector was designed to insert a 2A::iCreER::frt-neo-frt cassette into the translational stop site of the FBJ osteosarcoma oncogene locus (Fos) on chromosome 12. Apr 1, 2003 · Inducible transgenesis provides a valuable technique for the analysis of gene function in vivo. pA, a polyA Feb 20, 2022 · human estrogen receptor (called Cre-ERT2). We report the generation and characterization of mouse lines carrying glia lineage-specific transgenes expressing an improved variant of the tamoxifen-inducible Cre recombinase, CreERT2, where the recombinase is fused to a mutated ligand binding domain Jan 25, 2014 · The aim of the present study was to investigate the role of each estrogen receptors on the regulation of proteins involved with proliferation and differentiation of Sertoli cells from 15-day-old rats. However, acquired resistance to therapy has been noted in nearly one-third of women treated with tamoxifen and other endocrine therapies. 4–6 When Cre-ERT2 expressing Dec 17, 2024 · Mutation details: A targeting vector was designed to insert a CreERT2 fusion gene (Cre-ERT2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain), an SV40 polyA signal, and an FRT-flanked PGK-Neo cassette into the initiation codon of the cut-like homeobox 2 (Cux2) locus. We have analyzed the function of yeast and mammalian Hsp90 in regulating the ability of the human estrogen receptor (ER) to bind ligands in vivo and in vitro. Allele Symbol Allele Name Common Names Promoter Diseases Related to This Gene; Ddx4: DEAD box helicase 4: 13: Ddx4: endonuclease-induced mutation 1, University of Tsukuba Laboratory Animal Resource Center: ERT2: estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A) Oct 30, 2022 · Importantly, the synthetic ligand 4-hydroxy-tamoxifen (4-OHT, hereafter called tamoxifen) but not endogenous estrogens bind to the modified ERT2 [Citation 7]. A) Schematic map of SPC-Cre-ER T2 expression cassette. Oct 5, 2022 · Mutations in the estrogen receptor (ESR1) gene are common in ER-positive breast cancer patients who progress on endocrine therapies. CreER T2, Ert2-Cre, Gt(ROSA)26Sor tm1 Mutation details: A fusion protein consisting of cre recombinase and the mutated ligand binding domain of the human estrogen receptor (ERT2) was inserted into intron 1. , aromatase inhibitors, tamoxifen, and fulvestrant) are currently the standard of care. Oct 18, 2019 · By fusing the cytosol-localizing mutant estrogen receptor ligand binding domain (ERT2) to the N-terminal half of split Cre(2-59aa)-nMag, the TamPA-Cre protein ERT2-CreN-nMag is physically separated from its nuclear-localized binding partner, NLS-pMag-CreC(60-343aa). To rapidly induce RHOX10, we chose to employ the estrogen receptor T2 (ERT2) system, which drives swift translocation of the TF of interest to the nucleus in response to 4-hydroxytamoxifen of ligand-binding domain of the estrogen receptor (ERT2). As an example, mice expressing Cre-ERT2 can be crossed with another transgenic line containing a floxed “stop” signal preceding the sequence for a fluorescent reporter. 17 In Oct 18, 2024 · Rat Synapsin1 promoter, phage P1 Cre recombinase, human Estrogen receptor binding domain (ERT2) cDNA, SV40 poly A signal: Research application: Cre/loxP system: Specific Term and Conditions: In publishing the research results to be obtained by use of the BIOLOGICAL RESOURCE, an acknowledgment to the DEPOSITOR is requested. May 2, 1997 · The 90-kDa heat shock protein (hsp90) has been implicated in modulating steroid receptor function in vitro and in vivo. 5 kb of the first intron. Cre ERT2 LBD transgene is inserted in the Rosa26 (R26) locus, allowing ubiquitous expression. The mutant mouse estrogen receptor does not bind natural To trace PDGFRα-expressing cells in vivo, we generated a knock-in mouse line that expressed a fusion protein of green fluorescent protein (GFP), Cre recombinase (Cre), and mutated estrogen receptor ligand-binding domain (ERT2) under the control of the PDGFRα promoter. Without tamoxifen to drive nuclear localization of ERT2 Apr 1, 2022 · Moreover, Cre-activity is inducible since Cre is fused to mutated human estrogen receptor 1 (ER T2), which sequesters Cre from the nucleus until treating mice with tamoxifen (TAM, an exogenous ER agonist) (Fig. The promotor of the IEG Arc has been previously used to label neurons taking part in representations of experience Jun 1, 2017 · abbreviated as Cre – ERT2), a fusion of a mutated estrogen. , 2012) but no significant difference in proliferating progenitors of the SVZ was found (Zhu Aug 22, 2024 · To introduce temporal control in genetic experiments targeting the endothelium, we established a mouse line expressing tamoxifen-inducible Cre-recombinase (Cre-ERT2) under the regulation of the vascular endothelial cadherin promoter (VECad). In this study, we developed a novel SPC-Cre-ERT2 mouse model which can be applied for gene knock-out in Mar 1, 2017 · Mating of SPC–Cre-ERT2 knockin mice and EML4-ALK transgenic mice was performed to generate SPC–Cre-ERT2/EML4-ALK transgenic mice. 3-kb mouse Col1a1, collagen, type I, alpha 1, promoter. A DNA fragment containing transcription termination signals flanked by loxP sites is inserted between the promoter and the transgene, thereby preventing its transcription. 129S-Tg(UBC-cre/ERT2)1Ejb/J (hereafter abbreviated as Cre-ERT2), a fusion of a mutated estrogen receptor and Cre recombinase, was engineered to be more efficient and specific than the original Cre Inducible Cre-ERT recombinase technology is widely used for gene targeting studies. This proof of concept had many technical issues that were overcome over the years. Helices A & E are involved in the formation of the recombinase tetramer with the C terminus region of helix E known to form contacts with the C Mar 24, 2014 · Reversibly switchable proteins are powerful tools with which to explore protein function in vitro and in vivo. We report here the construction and characterization of a series of chimeric recombinases, each consisting of Cre fused to a mutated human oestrogen receptor (ER) ligand Apr 12, 2023 · (c) Cre recombinase is fused to a modified ligand-binding domain of the estrogen receptor (ERT2 LBD). 1999). Cre also eliminates the loxP-stop-loxP cassette, permitting widespread expression of Flpo-ERT2. Dec 17, 2024 · Mutation details: A targeting vector was designed to replace exon 2 of the chemokine (C-X3-C) receptor 1 (Cx3cr1) gene with a cre/ERT2 (cre recombinase fused to an estrogen receptor ligand binding domain) coding sequence, followed by an internal ribosome entry site (IRES) and an enhanced yellow fluorescent protein (EYFP). Cre also eliminates the loxP-stop-loxP cassette, permitting wide-spread expression of Flpo-ERT2. 많은 논문들을 보면 Cre에 estrogen receptor를 붙여서 사용하던데 그런 이유가 정확히 뭔가요 ㅠㅠㅠ 찾아보니 Cre를 핵안으로 더욱 효율적으로 이동 시켜주고 recombination을 잘되게 해주는것같은데 정확히 어떤 원리인지가 궁금하네요 Sep 27, 2018 · The Nfatc1 tm1. Jan 20, 2025 · Ordering Information; Donor DNA: mouse Crx promoter, phage P1 Cre recombinase gene, SV40 polyA signal, human Estrogen receptor binding domain (ERT2) Research application: Cre/loxP system: Specific Term and Conditions: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following We use cookies to personalize our website and to analyze web traffic to improve the user experience. Each Thy1 promoter region is Feb 12, 2021 · is fused with a mutated form of the estrogen receptor ligand binding domain (Cre/ERT2)/ERT2, in which Cre recombinase activity must be actively induced by tamoxifen administration. Flp-mediated recombination removed an FRT-flanked Lactb gene. In this study, we interrogated the intersect between supraspinal TNFR1 mediated neuronal signaling and sex specificity by selectively removing TNFR1 in Nex + neurons in adult mice (NexCre ERT2::TNFR1 f/f). Upon binding to 17beta-estradiol or related ligands, the encoded protein forms homo- or 007001 B6. In Cre-ERT2 mouse models, Cre recombinase is still expressed according to its driving regulatory sequences, but remains sequestered in the cytoplasm in complex with Hsp90. 29:494 , it was discovered that the transgene integrated Prion diseases are fatal neurodegenerative disorders caused by misfolding of the prion protein in the brain. [13] It has been Dec 20, 2024 · mouse Sox9 genomic DNA, EMCV internal ribosomal entry site (ires), bacteriophage P1 cre recombinase gene, human estrogen receptor binding domain (ERT2), mouse phosphoglycerate kinase promoter (PGK promoter), E. Cre-F primer binding sites, 561–579 bp of Sep 1, 2023 · TAM is a selective estrogen receptor modulator (SERM). Epub 2013 Jun 3. Mutation details: The transgene was designed with a creERT2 fusion gene, two copies of the mouse Thy1 promoter, and an enhanced yellow fluorescent protein (EYFP) cDNA sequence. . Hormone-activated Apr 14, 2016 · Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Cre/lox Apr 14, 2016 · Mutant Estrogen Receptor (ERT2) ligand-binding domain fusions with Cre recombinase are a key tool for spatio-temporally controlled genetic recombination with the Middle entry vector containing tamoxifen-inducible estrogen receptor (ERT2) fused to both ends of CRE recombinase for tamoxifen-inducible Cre recombination Apr 13, 2017 · The second generation of inducible Cre–ERT recombinase, hemizygous B6. The estrogen receptor contains 3 mutations (G400V, M543A, L544A) that increase sensitivity to tamoxifen. [3] Compared to pure ER Dec 20, 2024 · Ordering Information; Donor DNA: mouse Crx promoter, phage P1 Cre recombinase gene, SV40 polyA signal, human Estrogen receptor binding domain (ERT2) Research application: Cre/loxP system: Specific Term and Conditions: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following Aug 28, 1997 · Ligand-dependent chimeric Cre recombinases are powerful tools to induce specific DNA rearrangements in cultured cells and in mice. Allele Symbol Allele Name Common Names Promoter Diseases Related to This Gene; ERT2: estrogen receptor (ER), human ER with the triple mutation G400V/M543A/L544A) 5: ERT2: gastrointestinal stromal tumor()idiopathic hypereosinophilic syndrome()polyps, multiple and recurrent inflammatory fibroid, gastrointestinal() Apr 7, 2014 · The estrogen receptor (ER) and aryl hydrocarbon receptor (AhR) are ligand-activated transcription factors involved in estrogen or xenobiotic exposure, whereas the 90-kDa heat shock protein (HSP90), which is a ubiquitously expressed molecular chaperone, is involved in the signal transduction process. By expressing Cre at specific times or locations, you can precisely control expression of your gene of interest. We have Dec 16, 2009 · Methodology/Principal Findings. ER binds to specific DNA sequences called estrogen response elements (EREs) with high affinity and transactivates gene ex The estrogen receptor alpha (ERα) is a nuclear transcription factor that is expressed in more than 70% of all breast cancers. An estrogen receptor variant (ERT2) fused to the GAL4 transcriptional activator rapidly Dec 20, 2024 · Gene; Gene Symbol Gene Name Chr. When entering the nucleus, Cre catalyzes site-specific recombination events between loxP sites. Jan 15, 2024 · HSACreERT2 mice6 and Pax7CreERT2(FAN) mice7 were maintained heterozygous as the driver genes were (partially) replaced by the CreERT2 coding sequence. To delete a gene in a certain tissue temporally, tissue-specific and tamoxifen-inducible Cre transgenic mice are Background The selective estrogen receptor modulator tamoxifen, in combination with the Cre-ERT2 fusion protein, has been one of the mainstream methods to induce genetic recombination and has Nov 3, 2020 · Using the promotor of the IEG Arc to label hippocampal CA1 neurons representing experience. 24 In brief, the Aug 28, 1997 · Ligand-dependent chimeric Cre recombinases are powerful tools to induce specific DNA rearrangements in cultured cells and in mice. We recommend you draw out the recombination events so that the oligonecleotide Cre/ERT2 was created by fusing Cre recombinase cDNA with a mutated version of the estrogen receptor ligand binding domain (ERT2), thus creating a specific receptor for the tamoxifen metabolite OHT. The amino terminal domain encompasses residues 20–129 and this domain contains 5 alpha helical segments linked by a series of short loops. HSPC-P, human surfactant protein C promoter; Cre-ER T2, Cre coding sequence fused with a tamoxifen-inducible estrogen receptor. 4. You may decline these cookies although certain areas of the site may not function without them. rvya rbgo jdfmekp nlxtu vmzry hrjlr rwt tqjzh ncwd rqljcax