... is a special type of site-specific recombination. The Cre protein is a site-specific DNA recombinase. It can catalyze the recombination of DNA between specific sites in a DNA molecule. These sites, known as loxP sequences, contain specific binding sites for Cre that surround a directional core sequence where recombination can occur. It is often used in the generation of knockout and conditional knockout animals. - Cre-Lox Recombination - AbVideo™ - Support - Abnova
Conditional gene targeting using the Cre/loxP system enables specific deletion of a gene in a tissue of interest. For application of Cre-mediated recombination in pigment cells, Cre expression has to be targeted to pigment cells in transgenic mice. So far, no pigment cell-specific Cre transgenic line has been reported and we present and discuss our first results on use of Cre recombinase in pigment cells. A construct was generated where Cre recombinase is controlled by the promoter of the mouse dopachrome tautomerase (Dct) gene. The construct was functionally tested in vitro and introduced into mice. Following breeding to two reporter mouse strains, we detected Cre recombinase activity in telencephalon, melanoblasts, and retinal pigment epithelium (RPE). Our data demonstrate the feasibility of pigment cell-specific Cre/loxP-mediated recombination. Guyonneau, L.; Rossier, A.; Richard, C.; Hummler, E.; Beermann, F.
TY - JOUR. T1 - Dlx5/6-Enhancer Directed Expression of Cre Recombinase in the Pharyngeal Arches and Brain. AU - Ruest, Louis Bruno. AU - Hammer, Robert E. AU - Yanagisawa, Masashi. AU - Clouthier, David E.. PY - 2003/12. Y1 - 2003/12. N2 - Dlx5 and Dlx6, two members of the Distalless gene family, are required for development of numerous tissues during embryogenesis, including facial and limb development. This gene pair is expressed in tandem, transcribed toward each other and separated by a short intergenic region containing multiple putative enhancers. Targeted inactivation of Dlx5 and Dlx6 in mice results in multiple developmental defects in craniofacial and limb structures, suggesting that these genes are crucial for aspects of both neural crest and nonneural crest development. To further investigate potential developmental roles of Dlx5 and Dlx6, we used one of the Dlx5/6 intergenic enhancers to drive Cre recombinase expression in transgenic mice. Crossing Dlx5/6-Cre transgenic mice with ...
The Cre/ loxP site-specific recombination system has been used successfully for genome manipulation in a wide range of species. However, in Drosophila melanogaster, a major model organism for genetic
Excellgen Lambda Phage Integrase, Int [EG-40] - Description E coli lambda phage integrase Int is a site-specific tyrosine recombinase which mediates inserts and excises the phage genome into and out of the Escherichia coli chromosome. Lambda phage integrase has been used for LR and BP reactions in Gateway Cloning. Applications in vitro recombination assay Size 100 µg Source E coli overexpressing lambda phage Int
Development of inducible genetic switches for in vivo use with transgenic mice has revolutionized many areas in modern molecular biology. Combining two techniques, Cre/loxP-based genetic recombination and ligand-dependent activation of a chimeric protein, we generated transgenic mice which allow for the spatiotemporal control of expression and of activity of the proto-oncogene c-myc. To these ends, the gene encoding the tamoxifen-inducible c-mycER(T) fusion protein (mycER(T)) was inserted in the ubiquitously active ROSA 26 gene locus by gene targeting. In the resulting ROSAMER allele, generalized transcription of the mycER(T) gene is prevented by a preceding transcriptional stop sequence which is flanked by loxP sites. Crosses of ROSAMER transgenic mice with Mox2 cre transgenic mice revealed tight control of mycER(T) transcription in various tissues unless the transcriptional stop sequence was removed by cre-mediated excision. Furthermore, we were able to demonstrate tamoxifen-dependent activation of
Transgenic mice express Cre recombinase under the control of the mouse wingless-related MMTV integration site 7A protein (|i|Wnt7a|/I|). Cre recombinase expression is found in uterine luminal and glandular epithelial cells as well as other developmental processes. When used with a |i|loxP|/i|-flanked estrogen receptor allele, this mutant mouse strain may be useful in studies of female fertility and estrogen receptor function in uterine tissue.
Targeting posttraumatic inflammation is crucial for improving locomotor function. SIRT1 has been shown to play a critical role in disease processes such as hepatic inflammation, rheumatoid arthritis and acute lung inflammation by regulating inflammation. However, the role of SIRT1 in spinal cord injury (SCI) is unknown. We hypothesized that SIRT1 plays an important role in improving locomotor function after SCI by regulating neuroinflammation. In this study, we investigated the effect of SIRT1 in SCI using pharmacological intervention (SRT1720) and the Mx1-Cre/loxP recombination system to knock out target genes. First, we found that SIRT1 expression at the injured lesion site of wild-type (WT) mice (C57BL/6) decreased 4 h after SCI and lasted for 3 d. Moreover, administration of SRT1720, an agonist of SIRT1, to WT mice significantly improved functional recovery for up to 28 d post-injury by reducing the levels of pro-inflammatory cytokines, the number of M1 macrophages, the number of ...
We have used an aggrecan gene enhancer to generate a transgenic murine line (Acan-CreER-Ires-Luc) expressing firefly luciferase and tamoxifen activatable Cre recombinase (Cre-ER(T2) ). The expression and efficiency of the inducible Cre recombinase activity were tested in double transgenic mice created by crossing the Acan-CreER-Ires-Luc line with a Rosa26-lacZ reporter mouse. The expression pattern of the transgene of our line was restricted to cartilage from embryonic to adult stages. β-galactosidase staining was observed in growth plate, articular cartilage, as well as fibrocartilage of meniscus, trachea, and intervertebral discs. Similar staining was observed in a previously described Agc1 (tm(IRES-creERT2)) murine line. The presence of luciferase in our transgene allows the visualization of the transgene expression in live animals. Weekly measurements from 2 to 8 weeks of age showed a reduction in luminescence in knee joints between 2 and 4 weeks of age, but stabilization thereafter. Following the
Just to make things clear. This system does not need any less or any more PCR cloning then any other cre-lox system. You still need to insert your gene of interest into your donor vector. How you do it is up to you. You may have to use PCR to amplify your gene before ligating it into your donor vector. You may be able to cut said gene from an existing plasmid and ligate it into Multiple cloning site in the donor vector. Or if you are luck and willing to spend money, be able to find your gene already in a donor vector in libraries that use the CreatorTM vectors ...
The cellular mechanisms underlying feedback signaling from horizontal cells to photoreceptors, important for formation of receptive field surrounds of early visual neurons, remain unsettled. Mammalian horizontal cells express a complement of synaptic proteins that are necessary and sufficient for calcium-dependent exocytosis of inhibitory neurotransmitters at their contacts with photoreceptor terminals, suggesting that they are capable of releasing GABA via vesicular release. To test if horizontal cell vesicular release is involved in feedback signaling, we perturbed inhibitory neurotransmission in these cells by targeted deletion of the vesicular GABA transporter (VGAT), the protein responsible for uptake of inhibitory transmitter by synaptic vesicles. To manipulate horizontal cells selectively, an iCre mouse line with Cre recombinase expression controlled by connexin57 (Cx57) regulatory elements was generated. In Cx57-iCre mouse retina, only horizontal cells expressed Cre protein and its ...
About 5-10% of human gastric tumors harbor oncogenic mutations in the KRAS pathway, but their presence alone is often insufficient for inducing gastric tumorigenesis, suggesting a requirement for additional mutagenic events or microenvironmental stimuli including inflammation. Assessing the contribution of such events in preclinical mouse models requires Cre-recombinase-mediated conditional gene expression in stem or progenitor cells of normal and transformed gastric epithelium. We therefore constructed a bacterial artificial chromosome containing transgene (Tg) comprising the regulatory elements of the trefoil factor 1 (Tff1) gene and the Tamoxifen-inducible Cre recombinase (CreERT2) coding sequence. The resulting Tg(Tff1-CreERT2) mice were crossed with mice harboring conditional oncogenic mutations in Kras or Braf. Administration of tamoxifen to the resulting adult Tg(Tff1-CreERT2);KrasLSL-G12D/+ and Tg(Tff1-CreERT2);BrafV600E/+ mice resulted in gastric metaplasia, inflammation, and adenoma ...
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Tissue-specific gene inactivation using the Cre-loxP system has become an important tool to unravel functions of genes when the conventional null mutation is lethal. We report here the generation of a transgenic mouse line expressing Cre recombinase in endothelial cells. In order to avoid the production and screening of multiple transgenic lines we used embryonic stem cell and embryoid body technology to identify recombinant embryonic stem cell clones with high, endothelial-specific Cre activity. One embryonic stem cell clone that showed high Cre activity in endothelial cells was used to generate germline chimeras. The in vivo efficiency and specificity of the transgenic Cre was analysed by intercrossing the tie-1-Cre line with the ROSA26R reporter mice. At initial stages of vascular formation (E8-9), LacZ staining was detected in almost all cells of the forming vasculature. Between E10 and birth, LacZ activity was detected in most endothelial cells within the embryo and of extra-embryonic ...
Although the studies discussed above have shed light on the in vivo development and homeostasis of the mononuclear phagocyte system, mechanisms that control replacement of many macrophage subsets and resident cells remain mysterious. Most organs contain resident tissue macrophages, many of which have long half-lives. After irradiation and other forms of tissue injury, bone marrow-derived cells frequently repopulate the injured organs. Whether bone marrow-derived cells under inflammatory conditions assume the same phenotype and function as resident cells that populate tissues under steady state conditions remains largely unclear. Fate mapping experiments investigating steady state developmental potential will therefore have to avoid experimental inflammation. Toward this end, Cre recombinase expression under the control of promoters for key markers (such as lysozyme) have been used to activate conditional reporter genes so that all downstream daughter cells express a stable marker (9). It has to ...
Transcriptome profiling is widely used to infer functional states of specific cell types, as well as their responses to stimuli, to define contributions to physiology and pathophysiology. Focusing on microglia, the brains macrophages, we report here a side-by-side comparison of classical cell-sorting-based transcriptome sequencing and the RiboTag method, which avoids cell retrieval from tissue context and yields translatome sequencing information. Conventional whole-cell microglial transcriptomes were found to be significantly tainted by artifacts introduced by tissue dissociation, cargo contamination and transcripts sequestered from ribosomes. Conversely, our data highlight the added value of RiboTag profiling for assessing the lineage accuracy of Cre recombinase expression in transgenic mice. Collectively, this study indicates method-based biases, reveals observer effects and establishes RiboTag-based translatome profiling as a valuable complement to standard sorting-based profiling strategies.
Since the proposal of the classical model of the BG (3), substantial efforts have been made to uncover the selective contributions of the direct and indirect pathways to behavior. However, progress has been limited by the inability to access these neuronal populations due to the fact that they are anatomically intermixed. Here we used a genetic approach to dissect the function of these pathways by conditionally deleting the key striatal phosphoprotein, DARPP-32, in striatonigral and striatopallidal pathway neurons, using the D1R and D2R promoters to drive cell type-specific Cre recombinase expression (12). DARPP-32 plays an essential role in integrating signals from a number of behaviorally important neurotransmitters and neuromodulators that target the striatum (24). Thus, a loss of this protein would be expected to result in loss of function in each neuronal population. Supporting this, we found that deletion of DARPP-32 abolishes a key functional property of MSNs, corticostriatal LTP, in ...
Building on the basic strategy above, we describe methods for labeling inputs to specific cell types by considering the case in which starter cells are accessed using cell type-specific Cre driver lines. This is the most widespread approach and this example illustrates mechanisms underlying trade-offs between selectivity and efficiency; similar considerations apply to other approaches.. Numerous mouse lines are available that express Cre recombinase selectively in cell types of interest. To label the direct monosynaptic inputs to Cre+ neurons, Cre-dependent expression of G and TVA first need to be generated in the desired starter cell population (Fig. 2). This has been done by either infecting neurons in the desired starter cell location with a Cre-dependent helper virus (Figure 2; Wall et al., 2010; among many other publications cited previously), or by crossing the Cre driver line with a responder mouse line that has Cre-dependent expression of TVA and G (Li et al., 2013; Takatoh et al., 2013; ...
The advent of site-specific recombinase (SSR) technology and the Cre/lox system has led to numerous advances in molecular biology, and...
The Cre-lox system is a tool that involves the splicing of a specific pair of DNA sequences called lox sites with an enzyme called Cre recombinase. We implicated this system in attempts to manipulate gene expression that mimics randomly "selecting objects" to fill a knapsack in order to solve the knapsack problem. We needed a set of variant lox sites in order to create constructs that would yield different subsets of survival and fluorescence to optimally fill the knapsack. Initially the design to address the knapsack problem had several vital components using modules that consisted of lox sites that floxed the TetA gene and RFP. In our attempts to solve the knapsack problem, we assembled 10 new lox sites with mutations in the 8 bp region and floxed 16 out of 21 possible lox site combinations with red fluorescent protein with these variant lox sites. We built a construct with a fluorescent protein gene downstream of TetA to test for the presence of a terminator within the TetA gene. Along the ...
The Cre-lox system is a tool that involves the splicing of a specific pair of DNA sequences called lox sites with an enzyme called Cre recombinase. We implicated this system in attempts to manipulate gene expression that mimics randomly "selecting objects" to fill a knapsack in order to solve the knapsack problem. To do this we needed a set of variant lox sites in order to create constructs that would yield different subsets of survival and fluorescence to optimally fill the knapsack. Initially the design to address the knapsack problem had several vital components using modules that consisted of lox sites that floxed the TetA gene and RFP. In our attempts to solve the knapsack problem, we assembled 10 new lox sites with mutations in the 8 bp region and floxed 16 out of 21 possible lox site combinations with red fluorescent protein with these variant lox sites. We built a construct with a fluorescent protein gene downstream of TetA to test for the presence of a terminator within the TetA gene. ...
The Cre/lox system constitutes a method for the site-directed recombination of nucleic acids, in which the enzyme Cre-recombinase achieves recombination of DNA at specific target sequences, the so-called loxP-sites. In this process the activity of the Cre-recombinase is dependent on the control of a cell-specific promotor. This method presently is applied in great variety for systematic mutagenesis, for example for the generation of gene-knockouts in mice-models ...
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XerC is a site-specific recombinase of the bacteriophage lambda integrase family that is encoded by xerC at 3700 kbp on the genetic map of Escherichia coli. The protein was originally identified through its role in converting multimers of plasmid ColE1 to monomers; only monomers are stably inherited …
Introduction of new antibiotic resistance genes in the plasmids of interest is a frequent task in molecular cloning practice. Classical approaches involving digestion with restriction endonucleases and ligation are time-consuming. We have created a set of insertion vectors (pINS) carrying genes that provide resistance to various antibiotics (puromycin, blasticidin and G418) and containing a loxP site. Each vector (pINS-Puro, pINS-Blast or pINS-Neo) contains either a chloramphenicol or a kanamycin resistance gene and is unable to replicate in most E. coli strains as it contains a conditional R6Kγ replication origin. Introduction of the antibiotic resistance genes into the vector of interest is achieved by Cre-mediated recombination between the replication-incompetent pINS and a replication-competent target vector. The recombination mix is then transformed into E. coli and selected by the resistance marker (kanamycin or chloramphenicol) present in pINS, which allows to recover the recombinant plasmids
Conventional use of the site-specific recombinase Cre is a powerful technology in mouse, but almost absent in other vertebrate model organisms. In zebrafish, Cre-mediated recombination efficiency was previously very low. Here we show that using transposon-mediated transgenesis, Cre is in fact highly …
Fiber cells are among the longest lived cells in the body. Based on its expression pattern and putative antiapoptotic role in other systems, we hypothesized that Livin might contribute to lens cell longevity by suppressing cell death in the hypoxic core of the tissue. Alternatively, Livin, which is expressed by cells as they approach the OFZ (Fig. 7C), might have a role in organelle degradation. To test these hypotheses directly, we generated mice carrying a floxed Birc7 allele (see Supplementary Fig. S1), Birc7flox/flox mice were crossed with either LeCre16 or MLR1017 mice to conditionally delete Birc7 in the lens. The LeCre and MLR10 strains express Cre recombinase in lens and have been widely used to conditionally delete genes in this tissue. The two strains differ primarily in the timing of the onset of lens Cre expression (E8.75 for LeCre and E10.5 for MLR10). Since our data suggested that Livin expression does not commence until after E12.5 (Fig. 3B), we anticipated that the efficacy of ...
When these |i|Hif1a|sup|tm3Rsjo|/sup||/i| floxed mice are bred to mice that express Cre recombinase, resulting offspring will have exon 2 deleted in the |i|cre|/i|-expressing tissue(s). Mice from this strain can be crossed to strains expressing Cre recombinase in various tissues and may be useful for studies of the role of HIF transcription factors in von Hippel-Landau syndrome, adult erythropoiesis, inflammation, mammary epithelium, tumor angiogenesis, and lung development as examples.
Is it possible to get the striatum astrocyte-specific Cre recombinase-mediated knockout of XX gene knockout mice? Any one understand how to make the knockout locate in striatum or any other good ideas to knockout one gene in one special cell from one..
We also use the Cre-loxP system, in which a gene of interest is engineered to contain loxP sites flanking a critical region of the gene. A mouse containing the "floxed" gene is normal, because the loxP sites are silent. Upon expression of the Cre recombinase, which removes DNA sequences flanked by loxP sites, that gene is inactivated. We use three methods for inducing Cre in a region-specific manner in brain. In one, we breed mice containing a floxed gene to mice in which Cre is inducibly expressed under the tetracycline system described above. In the second, we breed them to mice that express modified forms of Cre, which can be activated upon local injection of a chemical in brain. In the third, we use viral vectors encoding Cre to create localized knockouts of the floxed gene. Together, these various approaches enable us to exert powerful control over a drug- or stress-regulated protein ...
Cre Recombinase Antibody is an affinity-purified Anti-Cre rabbit antibody that can be used in Western blot or IHC detection of Cre recombinase
Cre Recombinase Antibody is an affinity-purified Anti-Cre rabbit antibody that can be used in Western blot or IHC detection of Cre recombinase
Excellgen Cre Recombinase Exosome Like Vesicles [EG-1020] - Description Cre Recombinase exosome-like vesicles are 20 to 50 nm lipid vesicles isolated from cultured mammalian cells. These vesicles encapsulate high concentration of NLS-Cre recombinase, but do not contain virus and nucleic acids (plasmid DNA, RNA etc). Addition of 10 ~ 50 µl of the vesicles to reporter cells (such as
Emilin1 (E1) is a protein of the extracellular matrix that regulates TGFβ activity through proteolysis of the proTGFβ. E1 KO mice are hypertensive, with increased TGFβ activation. As E1 is expressed in blood vessels starting from embryonic life to adulthood, is still unknown whether the E1 KO phenotype results from a developmental defect or lack of a homeostatic role exerted in the adult. To dissect this issue, we inactivated E1 in smooth muscle cells (VSMCs) of adult mice, by the use of floxed E1 and CreERT2 [a tamoxifen (TAM) inducible Cre recombinase] under the control of the smooth muscle myosin heavy chain (Smmhc) promoter. When Smmhc-CreERT2 E1fl/fl mice were given TAM, blood pressure significantly increased (124±1 vs basal condition 106±1 mmHg) as well as myogenic response in resistance arteries (16.3±0.7 vs basal condition 11.4±0.1 % at 125 mmHg).. In order to evaluate the relevance of our findings in the human pathology, we enrolled 20 hypertensive and 20 normotensive patients ...
As described shortly after its discovery, CT-1 promotes cardiac myocyte hypertrophy by directing sarcomere assembly in series.3 At the ventricular structural level, in-series sarcomeric assembly leads to "eccentric" hypertrophy and chamber dilatation. Thus, increased myocardial expression of CT-1 immediately became a candidate for the molecular basis of pathological hypertrophy and remodeling in dilated cardiomyopathy phenotypes. The cardiac myocyte protective effects were also appreciated early in the investigation of CT-1s biological properties.4 Mice with genetic ablation of gp130 have hypoplastic hearts,5 which suggests a role for CT-1 or other IL-6-type cytokines in normal cardiac development. However, mice with ventricular myocyte-restricted knockout of gp130 using a ventricular myosin light chain 2 promoter that drives a Cre/lox recombination/knockout system from early in development have no cardiac abnormalities at birth,6 indicating that the hypoplastic heart phenotype of generalized ...
Fstl3floxflox were crossed with ? myosin heavy GDC0449 chain Cre transgenic mice that are maintained on C57BL6J background. Four different primer pairs were used for genotyping PCR. The loxP site in intron 2 was detected by using Primer1, SJL954 and Primer 2, SJL955 which amplify a 390 bp fragment for loxP site, while the Fstl3 wild type allele gives a 330 bp fragment. The loxP site in intron 5 was detected by using Primer 3, SJL956 and Primer 4, SJL986, which amplify an approximate 310 bp fragment for loxP site and a 270 bp fragment for wild type allele. Recombination by Cre leads to an allele that lacks exons 3, 4 and 5 of Fstl3 gene is detected by using primer pair of 1 and 4 that gives a 357 bp fragment. The ? MHC Cre transgene is detected by using the primer pair of 5 and, that amplifies a 300 bp fragment. Data are presented as meanSEM. Group differences were analyzed by two tailed Students t test or ANOVA ...
Full realization of the value of the loxP-flanked alleles generated by the International Knockout Mouse Consortium will require a large set of well-characterized cre-driver lines. However, many cre driver lines display excision activity beyond the intended tissue or cell type, and these data are frequently unavailable to the potential user. Here we describe a high-throughput pipeline to extend characterization of cre driver lines to document excision activity in a wide range of tissues at multiple time points and disseminate these data to the scientific community. Our results show that the majority of cre strains exhibit some degree of unreported recombinase activity. In addition, we observe frequent mosaicism, inconsistent activity and parent-of-origin effects. Together, these results highlight the importance of deep characterization of cre strains, and provide the scientific community with a critical resource for cre strain information.
Title. The function of inflammatory signalling pathways in acute and chronic liver disease and liver cancer. Concept. The aim of this proposal is to examine the role of inflammatory signalling pathways in murine models of liver and biliary disease by application of conditional gene targeting using cre/loxP technology. Previous studies have provided evidence that the NF-kB pathway and its activating kinase complex - consisting of three subunits: IKK1, IKK2 and NEMO - are crucial regulators of liver physiology and pathology, but their differential, cell specific functions in the liver are currently only poorly understood. The first part of this proposal will focus on the evaluation of molecular mechanisms underlying the development of hepatocellular carcinoma in a setting of chronic hepatitis. By using a novel mouse model of spontaneous liver cancer based on conditional deletion of NEMO in hepatocytes, the functions of cytokines, specific intracellular signalling pathways, the innate and adaptive ...
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MICER is a method developed by Allan Bradley. It consists of four sets of genomic clones that contain a loxP site in either orientation site and either the proximal or the distal half of a HPRT mini gene. After Cre mediated recombination between the loxP sites of two different MICER clones a complete HPRT mini gene is reconstituted and one can select for the Cre induced alteration of the genome. If the two loxP sites are located within the same chromosome and have the same orientation one will end up with a deletion of the sequences located between the loxP sites. If the loxP sites are inversely orientated Cre mediated recombination will induce an inversion of the sequences between the two loxP sites (it should be noticed though, that without selection prolonged presence of Cre can lead to a reversion of the fragment). If the loxP sites are situated on different chromosomes Cre mediated recombination will lead to reciprocal translocation between the two chromosomes, which is indeed the situation ...
The SLC30A8 locus encodes the Zn2+ transporter ZnT8, whose expression is largely restricted to α- and β-cells, endocrine cells of the pancreatic islet. Genome-wide association studies have revealed > 90 loci associated with Type 2 diabetes. The first such study identified a specific single nucleotide polymorphism that results in an amino acid substitution and a transporter with reduced activity. This risk variant is associated with increased Type 2 diabetes risk. More recently, rare loss-of-function variants were found to be protective. Here, we aimed to investigate the role of SLC30A8/ZnT8 in the regulation of glucagon secretion. ZnT8 was selectively deleted in the α-cell by crossing mice bearing floxed alleles at exon 1 with mice carrying a Cre recombinase transgene under the control of the preproglucagon promoter. Additionally, these mice were crossed to Rosa26 RFP mice for identification of α-cells. Fluorescence-activated sorting of RFP+ cells revealed that recombination at the RFP locus ...
Animals. Experiments were performed in 8- to 10-week-old male C57BL/6 mice obtained from The Jackson Laboratory. NSE-DTA mice (NSE-Stop-DTA mice) were donated by S. Itohara and Y. Kobayashi (RIKEN Center for Brain Science, Saitama Japan; refs. 34, 35) and were maintained through homozygous breeding pairs. Nestin-CreERT2/NSE-DTA mice were generated by crossing NSE-DTA mice with Nestin-CreERT2 mice (33) and then maintained through homozygous breeding pairs on a C57BL/6 background. In these transgenic mice (Nestin-CreERT2/NSE-DTA), tamoxifen-inducible Cre is expressed by NSCs under the nestin promoter and the loxP-STOP-loxP-IRES-DTA gene cassette is knocked into the NSE gene. After tamoxifen treatment, Cre recombinase deletes the STOP sequence in the NSC pool. Throughout maturation, the NSE promoter becomes active, inducing the expression of DTA, resulting in cellular programmed death. Thus, the generation of fully mature newborn granule neurons is dramatically decreased in these mice. The ...
Animals. Experiments were performed in 8- to 10-week-old male C57BL/6 mice obtained from The Jackson Laboratory. NSE-DTA mice (NSE-Stop-DTA mice) were donated by S. Itohara and Y. Kobayashi (RIKEN Center for Brain Science, Saitama Japan; refs. 34, 35) and were maintained through homozygous breeding pairs. Nestin-CreERT2/NSE-DTA mice were generated by crossing NSE-DTA mice with Nestin-CreERT2 mice (33) and then maintained through homozygous breeding pairs on a C57BL/6 background. In these transgenic mice (Nestin-CreERT2/NSE-DTA), tamoxifen-inducible Cre is expressed by NSCs under the nestin promoter and the loxP-STOP-loxP-IRES-DTA gene cassette is knocked into the NSE gene. After tamoxifen treatment, Cre recombinase deletes the STOP sequence in the NSC pool. Throughout maturation, the NSE promoter becomes active, inducing the expression of DTA, resulting in cellular programmed death. Thus, the generation of fully mature newborn granule neurons is dramatically decreased in these mice. The ...
Experimental approaches to evaluate cellular response to Potassium limitation. For this purpose, a series of isogenic strains derived from BY4741 and lacking one (TRK1 or TRK2) or both TRK genes encoding specific potassium uptake systems was constructed using the homologous recombination and Cre-lox system. ...
As noted above, a prominent feature of the dynamic regulation of FoxG1 is its upregulation in cells in the late multipolar phase prior to their migration into the cortical plate ( Figure 1A). To explore the significance of this upregulation, we have generated a Cre-dependent conditional loss-of-function allele of FoxG1 (FoxG1-C:Flpe, Figure S5) in order to allow us to. remove FoxG1 expression at specific stages of pyramidal cell migration. In constructing this conditional allele, the Flpe recombinase was inserted into the FoxG1 locus such that its expression is initiated upon removal of the loxP flanked FoxG1 gene ( Figure 4A scheme; Figure S5). Prior to Cre-mediated recombination, the expression of Flpe is attenuated by the FoxG1 coding and 3′UTR. domains, which act as a transcriptional stop cassette ( Dymecki and Kim, 2007, Joyner and Zervas, 2006, Luo et al., 2008 and Miyoshi Romidepsin and Fishell, 2006). By combining this conditional allele with a Flpe-dependent reporter line ...
J:160214 Maska EL, Cserjesi P, Hua LL, Garstka ME, Brody HM, Morikawa Y, A Tlx2-Cre mouse line uncovers essential roles for Hand1 in extraembryonic and lateral mesoderm. Genesis. 2010 May 12 ...
5crx_A mol:protein length:343 PROTEIN (BACTERIOPHAGE P1 CRE GENE) MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLSVCRSWAAWCKLN NRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPRPSDSNAVSLVMR RIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLGIAYNTLLRIAEI ARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERWISVSGVADDPNN YLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRYLAWSGHSARVGA ARDMARAGVSIPEIMQAGGWTNVNIVMNFIRNLDSETGAMVRLLEDGD ...
Floxed Ink4a/Arf Mouse - after deletion of the gene via crossing to a tissue-specific Cre line, mice can develop tumors, giving rise to various sarcomas, carcinomas, lymphomas, and metastatic melanoma
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Search for mouse SNPs represented in dbSNP by gene or genome region. Results include selected strains. Filter by dbSNP function class.