Drives sitespecific recombination at lox p or similar lox sites. Hence, advillin appears to be a promising candidate for the generation of a credriver mouse line with specificity in the pns ganglia. A blueprintfunded resource, the design, creation, and characterization of cre recombinaseexpressing credriver mouse lines on the c57b16 background. The offsprings usually contain a mixed c57bl6 and 129 genetic background carried from the. Charles gerfen, gensat offers transgenic baccre recombinase driver lines that allow for cellspecific gene manipulations in the central nervous system of the mouse. Here we report the generation and characterization of bactransgenic animals expressing cre recombinase under the regulatory elements of the advillin gene.
Progress in the somatosensory field has been restricted by the limited number of genetic tools available to study gene function in peripheral sensory neurons. Generation and analysis of an improved foxg1ires cre driver mouse line. A toolbox of credependent optogenetic transgenic mice for. The enzymes unique and specific recombination system is exploited to manipulate genes and chromosomes in a huge range of research, such as gene knock out or knock in studies. Use of musclespecific credriver lines of mice results in double transgenic offspring that express dux4fl mrna in skeletal muscles and develop an fshdlike myopathic phenotype. Here, we provide some most commonly used tissuespecific promoterenhancer of cre driver lines in table 1. Slc5a2 sweet pee enu mutagenized mouse model carrying a mutation on sglt2 gene which is a renal tubular sodiumglucose transporter.
The enzymes ability to operate efficiently in a wide range of cellular environments including. Nov 18, 2011 these will form the basis for selection of 250 new cre driver mouse lines that will be generated by the eu consortium eucommtools to be distributed worldwide through the european mutant mouse archive emma and by the canadian norcomm2ls project to be distributed by the canadian mouse mutant repository cmmr. The mouse lines are the latest addition to the cre driver network, an initiative to create a collection of mouse strains that offer scientists the ability to make precise alterations in brainrelated genes as a way of exploring their impact on brain function. Amyloid precursor protein, one of only 3 genes associated with inherited autosomaldominant ad mutations named for the location of families in which they were identified, i. Cre activity is restricted to adult striated muscle fibers and embryonic striated muscle. Mouse strains and cell lines available from the international mouse. Crossing flexdux4 mice with cre driver lines of mice leads to increased dux4fl mrna and protein expression. Generation and characterization of an advillincre driver. Dear colleagues, riken brc participates the create consortium as an advisor, the international coordination group. Tools for functional annotation of the mouse genome. Cre activity is restricted to adult striated muscle fibers and embryonic striated muscle cells of the somites and heart.
Cre mediated dna cleavage between two lox sites results in excision, integration, inversion, or translocation of the intervening sequence depending on the initial orientation and position of the lox p sites. Although a number of credriver mouse strains already exist for the kidney, development of additional strains with unique expression patterns is needed. Generation and characterization of an advillincre driver mouse line sandra zurborg, agnieszka piszczek, conception martinez, philip hublitz, mumna al banchaabouchi, pedro moreira, emerald perlas and paul a heppenstall abstract progress in the somatosensory field has been restricted by the limited number of genetic tools available to study. Here we report the generation and characterization of bactransgenic animals expressing crerecombinase under the regulatory elements of the advillin gene. Generation and analysis of an improved foxg1irescre driver. This study describes the generation of knockin mouse lines that express optogenetic activators or silencers in a cre recombinasedependent manner, and demonstrates the reliability and utility. Credriver mouse lines 3 current protocols in mouse biology volume 1 homogeneously validated cre lines, or individual assays can be applied to existing lines for which available information is incomplete andor requires con.
The jax cre repositorys aim is to provide the scientific community with a. The network is funded by the nih blueprint for neuroscience research. A various tissuespecific cre driver mouse lines have been generated to date, and new cre lines are still being developed. Double transgenic mice were then crossed with a cre driver line to generate mice in which gcamp6f expression is induced in the specific populations of neurons that express both cre and tta. These cre driver lines can be used to generate conditional mutations that are activated in distinct cell types, tissues or time points, or inducible mutations that are activated through administration of a drug. Thus, these mice should be of great use for investigating. The mouse lines are the latest addition to the cre driver network, an initiative to create a collection of mouse strains that offer scientists the ability to make. Cre recombinase is a tyrosine recombinase enzyme derived from the p1 bacteriophage. Welcome to the credriver network established by the nih neuroscience blueprint. In this overview, the minimum level of information that is ideally required to validate a cre. Nov 20, 2012 full realization of the value of the loxpflanked alleles generated by the international knockout mouse consortium will require a large set of wellcharacterized cre driver lines.
Mutant es cells eummcr european mouse mutant cell repository. The cre portal contains curated data about all recombinasecontaining transgenes and knockins developed in mice to provide a comprehensive resource delineating known recombinase activity patterns and allowing users to find relevant mouse resources for their studies. For the study of dopamine da neurons, promoter sequences driving expression of the. Cre transgenic mouse lines are available to generate somatic mutations, and thus the number of cre. Creloxp recombination system is a commonly used tool to achieve sitespecific genetic manipulation in genome. To this end, two transgenic mouse lines termed wnt1. Here we generated a credriver mouse line that expresses crerecombinase from the locus of the sensory neuron specific gene advillin. The eucommtools project created a new inducible cre driver mouse line resource that is available to researchers via infrafrontier repository. Although a number of cre driver mouse strains already exist for the kidney, development of additional strains with unique expression patterns is needed. Once produced, these transgenic lines must be validated for reliable, efficient, and specific cre expression and cre. Generation and characterization of an advillin cre driver.
Here we report the generation and validation of a tcf21pod1cre driver strain that expresses cre. These include mice where cre is under control of the locus of the voltagegated sodium channel na v 1. Generation and analysis of an improved foxg1irescre driver mouse line. Second row in a tg mouse that contains the gene xcre driver allele and the credependent reporter, transient cre activity in the embryo is permanently registered by the reporter allele. Transgenic credriver mouse lines are an important tool for optogenetic manipulations of neural circuit function.
Gene targeting restricted to mouse striated muscle lineage. Spatially and temporally regulated somatic mutations can be achieved by using the creloxp recombination system of bacteriophage p1. In order to develop gene knockouts restricted to striated muscle, we generated a transgenic mouse line expressing cre recombinase under the control of the human. Cre lox recombination involves the targeting of a specific sequence of dna and splicing it with the help of an enzyme called cre recombinase.
Promoters driving transgenic expression of the gene encoding cre recombinase are crucial to ensure selectivity in cremediated targeting of floxed alleles using the crelox system. These conditional gemms are then bred with one or several transgenic mouse lines expressing a site. In collaboration with the intramural program of the national institute of mental health pi. In the mouse, the cre lox system has been widely applied with a large number of cre driver transgenic mouse lines now available. Due to the usefulness of the creloxp system, numerous organspecific cre andor inducible cre transgenic mouse lines have been created and are still being developed. Here we report the generation and validation of a tcf21pod1 cre driver strain that expresses cre recombinase. Permits cellspecific killing of cells when crossed to appropriate credriver strain e. A blueprintfunded resource, the design, creation, and characterization of crerecombinaseexpressing cre driver mouse lines on the c57b16 background, which can be used to drive expression of reporter genes and conditionalready alleles in the mouse nervous system. Transgenic mouse lines are instrumental in our attempt to understand brain function.
Over the past few years many inducible andor tissuespecific cre mouse lines have been developed. Nih blueprint for neuroscience research has created several hundreds of cre driver mouse lines which are currently used by the worldwide neuroscience community. This resource contains over 220 cre driver mouse lines where tamoxifin inducible cre expression vectors have replaced the coding elements of genes with restricted tissue expression. As crerecombinase has no effect on dna except for those that contain loxp sites which are never found in mammalian dna, generating transgenic crerecombinase mouse models was a logical step in examining specific expression of genetic tools optogenetic, chemogenetic or other in neurons. A new cre driver mouse line, tcf21pod1cre, targets. Impccrelines international mouse phenotyping consortium. Several cre mouse lines are available that target subpopulations of neurons in the peripheral sensory system, in particular nociceptive neurons. A blueprintfunded resource, the design, creation, and characterization of crerecombinaseexpressing cre driver mouse lines on the c57b16 background, which can be used to drive expression of reporter genes and conditionalready alleles in.
Similarly, many credriver mouse lines are available for targeting neuronal subpopulations in the central nervous system. Conditional gene targeting in mice has provided great insight into the role of gene function in kidney development and disease. Specific expression is driven by the following cre driver lines. Finally, the more a given cre line is used, the more of these issues will be evaluated. Full realization of the value of the loxpflanked alleles generated by the international knockout mouse consortium will require a large set of wellcharacterized credriver lines. As cre recombinase has no effect on dna except for those that contain loxp sites which are never found in mammalian dna, generating transgenic cre recombinase mouse models was a logical step in examining specific expression of genetic tools optogenetic, chemogenetic or other in neurons. May 15, 2009 the generation of conditional knockout mice is a multiplestep process, which involves mating the flox mutant mouse line essential exons of the gene of interest are flanked by two loxp sites and the cre expressing mouse line. To systematically characterize whole brain gene expression patterns by in situ hybridization ish, cre driver mice from every line were crossed. Nih blueprint for neuroscience research has created several hundreds of cre driver mouse lines which are currently used by the worldwide neuroscience. Frontiers anatomical characterization of cre driver mice. A transgene was designed with the human doublecortin dcx promoterenhancer regions upstream of a creert2 fusion gene creert2. In this paper, we report a large and diverse set of new celltypespecific transgenic mouse lines, including 23 new driver lines 15 cre and 8 flp lines and 26 new reporter lines. This transgene was microinjected into the pronucleus of c57bl6j zygotes.
This cre ert2 transgenic mouse line was generated via lentitransgenesis. All eucommtool cre driver mouse lines are generated on a pure c57bl6n genetic background making this a unique resource for impc researchers. Mouse lines research baylor college of medicine houston. Charles gerfen, gensat offers transgenic bac cre recombinase driver lines that allow for cellspecific gene manipulations in the central nervous system of the mouse. Mouse models for neuroscience research welcome to the credriver network established by the nih neuroscience blueprint. Frontiers novel creexpressing mouse strains permitting. Jul 10, 2014 using this binary system, cellspecific transgene expression is accomplished practically through breeding of cre driver and reporter transgenic mouse lines, coinjection of cre driver and reporter engineered recombinant viruses into wild type mice, or the injection of recombinant viruses into transgenic mouse lines, which allows for additional. This creert2 transgenic mouse line was generated via lentitransgenesis. About nih neuroscience blueprint cre driver network. Most of the cre driver es cell lines are generated by the egfpcreer t2 knockin strategy. Crelox recombination involves the targeting of a specific sequence of dna and splicing it with the help of an enzyme called cre recombinase. The various organs were divided into 6 physiological systems.
Cre recombinase fused to a g400vm543al544a triple mutation of the human estrogen receptor ligand binding domain. To match the unmet need of a mouse model allowing specific and efficient in vivo genetic manipulation of cdc1, we generated two novel credriver lines, the xcr1 cre and karma cre mice, by knocking in an irescre expression cassette in the 3. This transgenic mouse line expresses cre initially in the midbrain and, after closure of the. Sep 11, 2011 similarly, many cre driver mouse lines are available for targeting neuronal subpopulations in the central nervous system. The goal of the project is to provide the neuroscience community with mouse strains that are suitable for the tissue and celltypespecific perturbation of gene function in the nervous system. Apr 19, 2018 apart from the creloxp system, another site. This, combined with other recombinases and transcriptional regulation systems dymecki et al. In addition, information regarding established cre. Molecular pain methodology open access generation and. By crossing with eucommeucommtools knockout mouse lines, the cre driver lines allow users to create conditional mutations that are activated in distinct cell types, tissues or time points, or mutations that are activated through administration of a drug. Generation and analysis of an improved foxg1irescre. Optimizing nervous systemspecific gene targeting with cre. Sep 11, 2011 hence, advillin appears to be a promising candidate for the generation of a cre driver mouse line with specificity in the pns ganglia.
New cre driver mouse lines experimental animal division. Heres how the different cre mouse model options enable. Read more conditional knockout models have greatly contributed to studies into gene function and disease processess by allowing the inactivation of gene function in specific tissues andor developmental timepoints in mice. A plethora of conditional mutants and creexpressing mouse lines is. A pancreas specific ptf1adriven cre mouse line causes. Cre loxp recombination system is a commonly used tool to achieve sitespecific genetic manipulation in genome. The generation of conditional knockout mice is a multiplestep process, which involves mating the flox mutant mouse line essential exons of the gene of interest are flanked by two loxp sites and the creexpressing mouse line.
To date, nih blueprint for neuroscience research cre projects have created several hundreds of cre driver mouse lines which are currently used by. A creinducible dux4 transgenic mouse model for investigating. Drives sitespecific recombination at loxp or similar lox sites. The system consists of a single enzyme, cre recombinase, that recombines a pair of short target sequences called the lox sequences. This system can be implemented without inserting any extra supporting proteins or sequences. For multiple cre driver mouse lines, parental transmissions of recombined flox alleles are reported. Cre expression is observed in astrocytes in the brain and spinal cord, as well as postnatal and adult gfapexpressing neural stem cells and their progeny in the brain. This project supports the generation of mice in which the expression of cre recombinase, a dna exchange enzyme, can be controlled temporally and spatially.
Mar 25, 2012 this study describes the generation of knockin mouse lines that express optogenetic activators or silencers in a cre recombinasedependent manner, and demonstrates the reliability and utility. A blueprintfunded resource, the design, creation, and characterization of crerecombinaseexpressing credriver mouse lines on the c57b16 background. Ptf1adriven cre lines are widely used to achieve genetic manipulation in a pancreas specific manner. The reporter lines express a variety of molecular tools that will enable a wide range of applications. Frontiers novel creexpressing mouse strains permitting to. A suite of transgenic driver and reporter mouse lines with. Transgenic cre driver mouse lines are an important tool for optogenetic manipulations of neural circuit function. Considerations when using credriver rodent lines for. The jax cre repositorys aim is to provide the scientific community with a centralized, comprehensive set of wellcharacterized cre driver lines and related information resources. The offsprings usually contain a mixed c57bl6 and 129 genetic background carried from the various cre and reporter lines. Use of musclespecific cre driver lines of mice results in double transgenic offspring that express dux4fl mrna in skeletal muscles and develop an fshdlike myopathic phenotype.
Conditional single vector crisprsacas9 viruses for efficient. Cre recombinase is a widely used tool in the field of molecular biology. To date, nih blueprint for neuroscience research cre projects have created several hundreds of cre driver mouse lines which are currently used by the worldwide neuroscience community. A plethora of conditional mutants and cre expressing mouse lines is now available to the research community that allows. Anatomical characterization of cre driver mice for neural. In the mouse, the most advanced mammalian genetic model organism, a binary system based on cre recombinase and its recognition sites lox sites has been applied in nearly all areas of brain research, with a large number of cre driver transgenic mouse lines now. Although this method revolutionized cell typespeci. Citeseerx document details isaac councill, lee giles, pradeep teregowda. The recombination pattern thus, neuron labeling is an integration. The development of smooth muscleexpressing credriver lines has allowed for exciting discoveries, including recent advances revealing the diversity of phenotypes derived from mature smc transdifferentiation in vivo using inducible creer t2 lines. Using this binary system, cellspecific transgene expression is accomplished practically through breeding of cre driver and reporter transgenic mouse lines, coinjection of cre driver and reporter engineered recombinant viruses into wild type mice, or the injection of recombinant viruses into transgenic mouse lines, which allows for additional. Mouse models have been an important tool for the study of smc functions.
A various tissuespecific credriver mouse lines have been generated to date, and new cre lines are still being developed. However, to date it has not been possible to generate cre drivers to target all subpopulations of peripheral sensory neurons without affecting other tissues in the organism. Cre recombinase activity is expected in white adipose tissue wat and brown adipose tissue bat transgenic cre activity, directed by the mouse aquaporin 2 promoter, is observed in kidney cells collecting duct, left and. Some of the artifacts and problems that can be associated with cre. Available strains include cre expressing strains, inducible cre strains, cre reporter strains, and loxpflanked floxed strains. Welcome to the cre driver network established by the nih neuroscience blueprint.