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| This transgenic floxed strain enables thiouracil (TU) tagging, a method developed for isolating RNA from specific cell types from within intact tissues without the need for cell dissociation. | |||||||||||||||
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Description
Strain Information
Former Names B6(D2)-Tg(CAG-HA/UPRT)985Code/J (Changed: 06-FEB-13 ) Type Mutant Strain; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System Noncarrier x Hemizygote (Female x Male) 21-MAR-13 Mating System Hemizygote x Noncarrier (Female x Male) 21-MAR-13 Species laboratory mouse Generation ?+N1 (31-MAY-13)
Generation DefinitionsDonating Investigator Chris Q Doe, University of Oregon, HHMI Description
Thiouracil (TU) tagging is a method developed for isolating RNA from specific cell types from within intact tissues without the need for cell dissociation.In this transgenic strain, a ubiquitous chicken β-actin-CMV (CAG) promoter drives expression of a loxP-GFP-3xstop-loxP cassette followed by a hemagglutinin (HA) epitope-tagged uracil phosphoribosyltransferase (UPRT) gene. Embryonic day 12.5 (E12.5) embryos and postnatal day 6 (P6) floxed animals show a widespread pattern of GFP fluorescence. The GFP-stop cassette effectively prevents any leaky read-through of UPRT expression from the floxed allele. Cre-induced excision of the floxed GFP-stop permits tissue-specific spatial expression of UPRT. Subsequent subcutaneous injection of 4-thiouracil (4TU) provides temporal specificity. Only UPRT+ cells exposed to 4TU produce thio-RNA which can then be purified for gene expression analysis via next-generation RNA sequencing (RNA-seq). TU tagging has been shown to have neglible effect on gene expression in cell lines and ubiquitous expression of UPRT has no effect on viability in mice.
When crossed with Tie2-cre (Tek-cre) animals, double-transgenic six-day-old mice show robust UPRT expression in PECAM+ (CD31) endothelial cells of the cerebellum and all other regions of the brain (e.g cortex, dentate gyrus, midbrain, choroid plexus, and hypothalamus) as well as Tie2-cre-expressing endothelial cells of the heart. Expression of UPRT is also seen in embryonic day 11.5 (E11.5) brain and heart.
When the floxed interval is excised by Math1-cre (Atoh-cre), mice show robust expression of UPRT in cerebellar granule neuron precursors (GNPs) of the postnatal day 6 brain.
Transplantation of UPRT+ cells can be carried out to create chimeric mice, useful in hematopoietic studies of cell migration and morphogenesis. TU tagging is complementary to cell isolation methods such as FACS panning and laser capture, with the added refinement of isolating only nascent RNA.
Development
A CAG promoter composed of chicken β-actin promoter and cytomegalovirus (CMV) early enhancer element drives expression of a loxP-GFP-3xSV40-loxP cassette followed by hemagglutinin (HA) epitope-tagged uracil phosphoribosyltransferase (UPRT) gene from Toxoplasma gondii. The transgenic vector was injected into B6D2F1 hybrid oocytes. Resultant mice were backcrossed to C57BL/6 for 4 generations by the donating laboratory.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| Considerations for Choosing Controls | ||
Related Strains
Strains carrying other alleles of GFP
View Strains carrying other alleles of GFP (317 strains)
009668 B6;D2-Tg(Actb-Uprt)372Cdoe/J
Phenotype
Phenotype Information
Currently there is no phenotype information for this strain.This mouse can be used to support research in many areas including:GFP related
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Tg(CAG-GFP,-Uprt)985Cdoe | ||
|---|---|---|---|
| Allele Name | transgene insertion 985, Chris Q Doe | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | CAGFPstopUPRT; | ||
| Strain of Origin | (C57BL/6 x DBA/2)F1 | ||
| Expressed Gene | GFP, Green Fluorescent Protein, jellyfish | ||
| Green Fluorescent Protein (GFP), derived from the jellyfish Aequorea victoria, is a versatile reporter molecule which has found use in many biological applications. In some constructs the original molecule has been modified in order to enhance its fluorescence intensity (EGFP, enhanced GFP). When utilized in a transgenic construct, tissue expressing sufficient amounts of GFP will fluoresce when exposed to a 488 nm light source. | |||
| Expressed Gene | Uprt, uracil phosphoribosyltransferase (FUR1) homolog (S. cerevisiae), mouse, laboratory | ||
| Promoter | CMV, cytomegalovirus, human | ||
| Molecular Note | A CAG promoter composed of chicken beta-actin promoter and cytomegalovirus (CMV) early enhancer element drives expression of a loxP-GFP-3xSV40-loxP cassette followed by hemagglutinin (HA) epitope-tagged Toxoplasma gondii uracil phosphoribosyltransferase (UPRT) gene. Line 985 was generated. [MGI Ref ID J:191564] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(CAG-GFP,-Uprt)985Cdoe,MELT
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
Gay L; Miller MR; Ventura PB; Devasthali V; Vue Z; Thompson HL; Temple S; Zong H; Cleary MD; Stankunas K; Doe CQ. 2013. Mouse TU tagging: a chemical/genetic intersectional method for purifying cell type-specific nascent RNA. Genes Dev 27(1):98-115. [PubMed: 23307870] [MGI Ref ID J:191564]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX18
Colony Maintenance
Breeding & Husbandry Homozygoes and hemizygotes are viable and fertile. Mating System Noncarrier x Hemizygote (Female x Male) 21-MAR-13 Hemizygote x Noncarrier (Female x Male) 21-MAR-13 Diet Information LabDiet® 5K52/5K67
Pricing and Purchasing
Pricing, Supply Level & Notes, Controls
| Pricing for USA, Canada and Mexico shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $232.00 Female or Male Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe
Price per Pair (US dollars $) Pair Genotype $302.00 Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe x Noncarrier $302.00 Noncarrier x Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
| Pricing for International shipping destinations |
|
Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $301.60 Female or Male Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe
Price per Pair (US dollars $) Pair Genotype $392.60 Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe x Noncarrier $392.60 Noncarrier x Hemizygous for Tg(CAG-GFP,-Uprt)985Cdoe Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
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Standard Supply
Repository-Live.
Repository-Live represents an exclusive set of over 1500 unique mouse models across a vast array of research areas. Breeding colonies provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. If a Repository strain is not immediately available, then within 2 to 3 business days, you will receive an estimated availability timeframe for your inquiry or order along with various delivery options. Repository strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping. We will note and try to accommodate requests for specific ages of Repository strains but cannot guarantee provision of these strains at specific ages. However, if cohorts of mice (5 or more of one gender) are needed at a specific age range for experiments, please let us know.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.Ordering Information
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