Inducible cre mice pdf merge

Inducible depletion of adult skeletal muscle stem cells. It is important to note that inducible crelox systems also allow one to limit unwanted cre activity and associated side effects for instance, ectopic recombination due to transient cre expression during development or potential toxic effects due to prolonged high levels of cre activity 7, 8. Inducible glomerular erythropoietin production in the adult. Podocytespecific expression of tamoxifeninducible cre. Tamoxifenindependent recombination in the ripcreer mouse. Cre expression in unexpected tissues including germline incomplete recombination efficiency mosaicism. Pdf inducible and reversible silencing of the pvalb gene in.

We employed mice with an inducible cre, and conditionally expressed dta to deplete or gfp to track scs. Inducible cre mice article pdf available in methods in molecular biology 530. Podocytespecific expression of tamoxifen inducible cre recombinase in mice nephrology dialysis transplantation, jul 2010 hideki yokoi, masato kasahara, masashi mukoyama, kiyoshi mori, koichiro kuwahara, junji fujikura, yuji arai, yoko saito, yoshihisa ogawa, takashige kuwabara, et al. Microglia promote learningdependent synapse formation through brainderived neurotrophic factor christopher n. Briefly, pancreata from male animals were dissected, fixed in formalin, embedded in paraffin and entirely sectioned at a thickness of 5 m. However, the research community requires more cre recombinase expressing transgenic mouse strains cre drivers that restrict expression to specific cell types. The purpose of this study was to create and characterize an inducible, skeletal musclespecific cre transgenic mouse strain. Endothelial tracing was achieved by tamoxifen administration to inducible tie2 cre mice tamoxifen inducible cre activation under the control of endotheliumspeci. The use of cre recombinase and loxp sites in genetically modified mouse models allows for tissue specific control over genetic modifications, enhancing our understanding of gene functions. Methodology open access inducible cre transgenic mouse strain for skeletal musclespecific gene targeting john j mccarthy1,2, ratchakrit srikuea2,5, tyler j kirby3, charlotte a peterson1,4 and karyn a esser1,2,6 abstract background.

Inducible activation of cre recombinase in adult mice causes gastric epithelial atrophy, metaplasia, and regenerative changes in the absence of. Simplifying the combined use of crisprcas9 and creloxp. Littman,5,7 and wenbiao gan1 1molecular neurobiology program, the kimmel center for biology and medicine at the skirball institute, department of. Tamoxifen administration allows for ablation of predetermined genes in. Pdf system for tamoxifeninducible expression of cre. Get supplemental information, a quote, and estimated timeframe for generating your inducible knockout mouse line. Verma, march 29, 2004 recently, several systems designed to trigger rna. Podocyte cellspecific expression of doxycycline inducible cre recombinase in mice. A method of gene targeting that allows the inducible inactivation of a target gene in mice is presented. Mice expressing gfp and creer in osteochondro progenitor.

Expression levels are dependent on the dose of the agent administered. These transgenic mice express a tamoxifen inducible cre recombinase driven by the mouse col1a2, collagen, type i, alpha 2, promoter. When crossed with a rosa26lacz reporter tamoxifen induced cre recombination is detected in 97%99% of adipocytes in white. An inducible cre system is controlled by cellspecific regulatory elements promoters and enhancers and temporally inducible way by exogenous inducer such as tamoxifen tam or tetracycline tet 1,10,1417. Cronin and colleagues 2001 used a modified version of the lac operatorrepressor system to control expression of an exogenously introduced reporter gene in mice. Inducible knockout mouse for timedependent gene control. Crisprcas9 inducible systems were also developed, which rely either on an inducible promoter or on the activation of a. Deletion was complete in liver and nearly complete in lymphocytes. Tamoxifen tm inducible cre recombinases are widely used to perform gene inactivation and lineage tracing studies in mice. The transgene is expressed in osteochondro progenitor cells in the developing limb buds and in a subpopulation of periosteal cells that is closely associated with the cortical bone. The cd4 cre construct was created by replacing the the lck proximal promoter of a construct used to make a lck cre transgenic mouse taconic line 4197 with the mouse cd4 promoterenhancersilencer, which was provided by dr. Essential role of developmentally activated hypoxiainducible. Betacell fraction and mass were assessed as previously described. We first verified that the crelox recombination occurred correctly in cartilage in col2creert.

The analysis of the role of genes in important brain functions like learning, memory and synaptic plasticity requires gene inactivation at the adult stage to exclude developmental effects, adaptive changes or even lethality. Adipocyte dynamics and reversible metabolic syndrome in. Elevated pth induces endothelialtochondrogenic transition. Inducible knockout of mouse zfhx3 emphasizes its key role. Inducible deletion of vhl in reninexpressing cells doxycycline inducible vhl knockout in reninexpressing cells reninicre vhlflfl mice was achieved by crossing vhlflfl mice with transgenic mice expressing a reverse tetracycline transactivator from the mouse renin gene locus mrenrttam2 mice and lc1 mice, which express cre. To generate the adipoqcreer bac transgenic mice, the 245 kb rp2390g21 bacterial artificial chromosome bac containing at least five mouse genes was modified by replacing the starting atg and 222 bp of the adiponectin gene with the starting atg and sequence encoding the cre ert, cre recombinase and estrogen receptor 1 mouse fusion gene. Nerves of the peripheral nervous system contain two classes of schwann cells myelinating schwann cells that ensheath large caliber axons in myelin, and remak schwann cells that surround smaller axons and do not myelinate. Podocyte cellspecific expression of doxycycline inducible. To do this, we generated mice with an inducible deletion of endothelial s1p 1r by crossing mice with floxed s1p 1r s1p 1r ff with mice that express a tamoxifen inducible form of cre recombinase icreert2 under the transcriptional control of the plateletderived growth factorb pdgfb gene. Conditional and induced gm models university of oulu.

Our chemicalgenetic approach will help to investigate when, where, and how genes control neuronal circuit dynamics, and organize, for example, sensory signal processing. Crelox basics generating knockout mice webinar 17mar2016. These experiments were performed in male adult mice. An inducible cre mouse line to sparsely target nervous. Tamoxifeninduced creloxp recombination is prolonged in. A timedependent conditional knockout mouse defines an inducible animal model in which a gene of interest is floxed thus temporally controllable at a given timepoint in embryonic, postnatal or adult animals. Cggtrosa26sortm1rtta,egfpnagyj 005670 dow 2014 plos one pmid. Transdifferentiation of glioblastoma cells into vascular. Cardiacspecific inducible and conditional gene targeting. Betacell mass and glucose homeostasis in double mutant gr overexpressing mice. Singleneuron labeling with inducible cre mediated knockout in transgenic mice paul young, li qiu, dongqing wang, shengli zhao, james gross, guoping feng supplemental figure 1. While tools exist for genetic targeting of myelinating schwann cells, such reagents have been challenging to generate for the remak population in part. Inducible activation of cre recombinase in adult mice.

These transgenic mice have a tamoxifen inducible cre mediated recombination system driven by the mouse plp1, proteolipid protein myelin 1 promoter. In order to achieve temporally controlled somatic mutagenesis, the cre loxprecombination system has been complemented with the tamoxifen inducible fusion protein. Springer nature is developing a new tool to find and evaluate protocols. Egr1, an immediate early gene, is undetectable in the embryonic nervous system, and is increasingly induced postnatally. Mar 22, 2005 conditional and inducible transgene expression in mice through the combinatorial use of cre mediated recombination and tetracycline induction gusztav belteki, 1 jody haigh, 1 nikolett kabacs, 1 katharina haigh, 1 karen sison, 1 frank costantini, 2 jeff whitsett, 3 susan e. Crelox technology and breeding schemes for research. Lentiviral transgenesis was achieved using transgenic oocytes carrying a creert2 an inducible form of cre transgene driven by a glial fibrillary acidic protein gfap promoter.

The ripcreer mouse tg ins2creesr1 1dam is among the few available. To study the roles of key transcription factor networks, growth factors, and signaling molecules in the endoderm, notochord, and floorplate, we developed an inducible cre expressing system for altering gene function in this tissue. Inducible cre transgenic mouse strain for skeletal musclespecific gene targeting john j mccarthy1,2, ratchakrit srikuea2,5, tyler j kirby3, charlotte a peterson1,4 and karyn a esser1,2,6 abstract background. Effective experimental models are characterized by low background levels of the regulated gene and induction to high levels with subphysiological levels of. This transgene expresses cre recombinase under the control of an inducible mx1 promoter, which is silent in healthy mice, and active in the liver and in lymphocytes after induction with ifn or pipc. Cd4cre transgenic taconic biosciences humanized mice. When these caggcreer tm mice are bred with mice carrying a floxed target gene, tamoxifen inducible cre mediated recombination results in deletion of the floxed sequences in widespread cells or tissues of the offspring, including developing embryos and cultured cells derived from transgenic mice.

The inducible and reversible gene expression that occurs with this bacterial system ensures that genes within the lac operon are expressed only when necessary. Background the inducible crelox system is a valuable tool to study gene function in a spatial and time restricted fashion in mouse models. Regulating rtta and target expression with cre teton krasg12d to model pancreatic cancer b6. Apr 01, 2019 to understand gene function, the cre loxp conditional system is the most powerful available for temporal and spatial control of expression in mouse. A common induction mechanism is the use of tamoxifendependent cre recombinase creer and creert2, but the effects of tamoxifen at doses normally used in lineagetracing studies on normal adult mammary gland homeostasis are not known.

Conditional and inducible transgene expression in mice. These mice which carry gfapcreert2 express creert2 in astrocytes and were generated by classical pronuclear injection feil et al. This transgene is 97%99% effective in inducing gene recombination. The use of the cre loxp system for gene targeting has been proven to be a powerful tool for. The use of the cre loxp system for gene targeting has been proven to be a powerful tool for understanding gene function. Using cre lox technology, we developed an inducible endothelialspecific prongf transgenic mouse prongf loxp that overexpresses gfpconjugated cleavageresistant prongf123 when crossed with vecadherincreert2 cre. The cd4 cre transgenic mouse was originally developed in the laboratory of dr. Jul 24, 2014 lineage tracing using inducible genetic labeling has emerged to be a powerful method for interrogating the developmental fate of cells in intact tissues.

Cre recombinaseinducible rna interference mediated by. Microglia promote learningdependent synapse formation. Inducible gene inactivation in neurons of the adult mouse. The cre lox sitespecific recombination system has emerged as an important tool for the generation of conditional somatic mouse mutants. Spccreert2 transgenic mouse for temporal gene deletion in. Expression of prongf, inflammatory mediators, ngf and vegf was evaluated by pcr, western blot and immunohistochemistry. To address these problems, a highthroughput method for largescale production that. Tamoxifen inducible cre system is achieved by modified cre protein fused with the estrogen receptor. To achieve skeletal musclespecific expression, the human. Heritable and inducible gene knockdown in astrocytes or. Hundreds of different cre mice are already available, with different patterns of cre expression allowing conditional knockout in different tissues. The teto cre mice also provided by dr andras nagy, carry the cre coding sequence. To determine the ability of cre recombinase to induce recombination specifically in bone of inducible cre mouse which has osteoblasts and odontoblastsspecific collagen promoter, inducible cre mouse was crossed with the rosa26 mouse with lacz reporter.

Cre recombinase inducible rna interference mediated by lentiviral vectors gustavo tiscornia, vinay tergaonkar, francesco galimi, and inder m. Basic cre lox mechanism strain types and breeding schemes o tissuespecific knockouts o general knockouts o inducible knockouts o reporters cre lox web resources finding mice, cre. Basic cre lox mechanism strain types and breeding schemes o tissuespecific knockouts o general knockouts o inducible knockouts o reporters cre lox web resources finding mice, cre activity data, and more 7. To permit temporallyspecific genetic manipulation, we utilized cre er t2, a tamoxifen inducible version of cre recombinase, and drove its expression with a 92 kb bac fragment covering early growth response 1 egr1. The method uses an interferonresponsive promoter to control the expression of cre recombinase. Stifter and melanie greter institute of experimental immunology, university of zurich, winterthurerstrasse 190, 8057, zurich, switzerland cre and creer mouse strains are powerful tools that have proven invaluable for investigating the function of genes and for the fatemapping of cell populations. Novel transgenic mice for inducible gene overexpression in. The influence of tamoxifen on normal mouse mammary gland.

This modified bac transgene was microinjected into. The simplest crelox system to generate a knockout ko mouse model has two components that need to be genetically engineered in the murine genome. The crelox sitespecific recombination system has emerged as an important tool for the generation of conditional somatic mouse mutants. Mhc extensively used to drive cardiomyocyterestricted recombination of floxed genes in animal models. Inducible cre transgenic mouse strain for skeletal muscle. Actinin cx43 yfp merge inset legend on next page 230 cell reports 8, 229241, july 10, 2014. Toassess the efficiency of induced cre mediateddeletion ofa target gene, wegenerated mice transgenic for a mxcreexpression vector 8 andcrossed themto a strain. Selective deletion of the endothelial sphingosine1phosphate. Hence, mice harboring a floxed target gene and a mx cre transgene acquire an inactivating mutation of the target gene upon treatment of the mice with ifn or pipc. Mice hemizygous for this adipoqcreert2 bac transgene are viable and fertile, with expression of a tamoxifen inducible cre recombinase cre ert2 directed to adipose tissue by the promoterregulatory regions of the mouse adiponectin adipoq locus on the bac transgene. Inducible overexpression of endothelial prongf as a mouse. A creinducible diphtheria toxin receptor mediates cell. The abcs of choosing a cre strain to generate ko mice.

Pu,2,3 chingpin chang,4 and bin zhou1,5,6 1department of genetics, albert einstein college of medicine of yeshiva university, bronx, new york 2department of cardiology, childrens hospital. Deletion was complete in liver and nearly complete in lymphocytes within a. Disruption of myelin in the corpus callosum of dttreated mogi cre idtr mice. Moon1,4,5 to study the roles of key transcription factor. The transgene insert contains a fusion product involving cre recombinase and a mutant form of the mouse estrogen receptor ligand binding domain.

Inducible glomerular erythropoietin production in the. When crossed with a strain containing a loxp site flanked sequence of interest, the offspring are useful for generating tamoxifeninduced, cre mediated targeted deletions. Singleneuron labeling with inducible cre mediated knockout in transgenic mice paul young1,4, li qiu1, dongqing wang1, shengli zhao1, james gross1,3, and guoping feng1,2 1department of neurobiology, duke university medical center, research drive, durham, north carolina 27710, usa 2department of pathology, duke university medical center, research drive, durham, north carolina 27710, usa 3duke. This method allows one to control gene activity in space and time in almost any tissue of the mouse, thus opening new avenues for studying gene function and for establishing sophisticated animal models of human diseases. Although community efforts are underway to fully characterize newly produced cre driver mice. Inducible gene knockout in mice article pdf available in journal of investigative dermatology 47. Here, cre was used to delete a segment of the dna polymerase beta gene flanked by ioxp recombinase recognition sites. When we used a commonly utilized tamoxifen dose 100 mgkgday. Optimizing tamoxifeninducible creloxp system to reduce.

Usingcx3cr1 creer todrivediphtheriatoxinreceptor expression in microglia, we found that microglia could be speci. Inducible gene targeting in mice using the cre lox system brian sauer laboratory of biochemistry and metabolism, national institute of diabetes, digestive and kidney disease, national institutes of health, building 10, room 9n119, bethesda, maryland 208921800 email. Inducible transgenic mouse models allow for the activation of genes in specific cells and tissues at specific times. Yfp fluorescence was imaged in sagittal sections from slickv transgenic mice. In this study, we developed a spc cre ert2 mouse model, in which a tamoxifen inducible cre recombinase cre ert2 is under the control of the human surfactant protein c spc promoter.

In this system, we utilize highperforming aba inducible cre, gib inducible flp, and rap inducible. Merge inset vecadherincrelacz vecadherincreyfp ctnt tie1crelacz tie1crelacz d tie1creyfp merge inset off on. Another technical concern regarding the use of cre mice is that these mice either can exhibit phenotypic abnormalities and. This strategy relies on the limited background activity of the modified cre recombinase creer in the absence of its inducer, the competitive estrogen receptor ligand, tamoxifen. Transgenic mice inducible gene knockout somatic mutagenesis creer recombinase tamoxifen in utero rosa26 r26r xgal staining mouse models of human disease. Functional study of gene using inducible cre system. Endothelial tracing was achieved by tamoxifen administration to inducible tie2cre mice tamoxifeninducible cre activation under the control of endotheliumspeci. Aug 27, 2015 therefore, the applicability of sc studies to attenuate muscle loss due to nmj deterioration as observed in neuromuscular diseases and aging is ambiguous. Although several cre loxpbased gene knockout mouse models have been generated for the study of gene function in alveolar epithelia in the lung, their applications are still limited. Creer mice expressing tamoxifeninducible cre recombinase that allow for speci. Nih public access transgenic mice nat neurosci single. Generation of aggrecancreert2 knockin mice for inducible cre. The use of the creloxp system for gene targeting has been proven to be a powerful tool for understanding gene function. We generated prx1creergfp transgenic mice that express tamoxifen inducible cre recombinase and gfp under the control of a 2.