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- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
- Terms of Use
Description
Strain Information
Former Names 129-Gt(ROSA)26Sortm1(Dsred2/EGFP)Luo/J (Changed: 31-OCT-06 ) Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Mating System Heterozygote x Heterozygote (Female x Male) Species laboratory mouse Generation N4+N1F1 (03-JAN-07) Donating Investigator Liqun Luo, Stanford University, HHMI Description
MADM-RG mice are viable with no gross behavioral or observable abnormalities. Homozygous females produce less pups to weaning age compared to heterozygotes. These mutants are designed for MADM (mosaic analysis with double markers), and must be crossed to mice harboring a reciprocal mutation at the same locus (see Stock No. 006041 or Stock No. 006075, MADM-GR (EGFP/Dsred2)). The resulting offspring have one copy of each reciprocal mutation on homologous chromosomes ("trans-heterozygous") and must next be bred to a Cre-expressing strain for fluorescent protein expression. Prior to Cre-recombination, double mutant mice do not have colored cells: the chimeric genes do not produce functional proteins because their coding sequences are interrupted by the beta-actin intron in different reading frames. After DNA replication (G2 phase) in double mutant mice, Cre-recombinase introduction that facilitates inter-chromosomal recombination aligns the respective N- and C-terminal coding sequences for each of the reporter genes on the same chromosome. The following chromatid segregation (X or Z) determines daughter cell phenotype (recombinant sister chromatids into the same daughter cell leads to double reporter expression [a G2-Z event], while independent segregation into separate daughter cells leads to expression of EGFP or Dsred2-MYC [a G2-X event]). If an additional targeted mutation of interest is introduced distal to the Gt(ROSA) locus, only homozygous cells will be singly labeled following G2 Cre-introduction. The homozygous mutant and wild type cells can then be distinguished by which single reporter they express. Reporter protein tissue specificity, expression levels, and frequency of recombination are thus determined by the promoter controlling Cre expression. Using this MADM system, a researcher can generate genetic mosaics in which an individual organism contains somatic cells of different genotypes. This allows the researcher to ascertain lineal relationships and pleiotropic gene function in multicellular organisms. These mice may also be useful in studies of cell differentiation and mitosis.Mice harboring this MADM-RG mutation are also available congenic on a C57BL/6J genetic background (see Stock No. 006080). A control strain (Stock No. 006053, MADM-GG (EGFP/EGFP)) is also provided in which EGFP is in-frame on the same chromosome. Other important features of the MADM system are listed below. Cre-recombinase introduction in cell phase G0 or G1 results in double reporter expression. While EGFP expression can be visualized in vivo and in fixed samples, Dsred2-MYC can not. Anti-MYC immunofluorescence allows simultaneous visualization of both reporter genes. Single copy Dsred2-MYC is inadequate for live visualization.
Development
A targeting vector was designed to contain the N-terminal portion of a red fluorescent protein (Dsred2; Clonetech), beta-globin intronic sequence (itself containing a loxP-flanked neomycin resistance gene), and C-terminal portion of a mutant enhanced green fluorescent protein (mut4-EGFP; Okada et al, Exp Neurol 1999 156:394-406). This construct (RG) is preceded by the cytomegalovirus beta-actin enhancer-promoter and followed by the SV40 T antigen poly A signal, allowing for ubiquitous and high-level expression of the reporter genes. This entire construct was inserted into the Gt(ROSA)26Sor locus via electroporation of (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were microinjected in C57BL/6 blastocysts and chimeric progeny were established. Mutant mice were bred to 129S1/SvImJ (Stock No. 002448) for at least three generations, and then intercrossed to homozygosity before arriving at The Jackson Laboratory.
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 002448 129S1/SvImJ | (approximate) | |
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (170 strains)
006080 B6.129-Gt(ROSA)26Sortm2Luo/J
Additional Web Information
Cre-lox Systems
Fluorescent Proteins/lacZ Systems
Phenotype
Phenotype Information
assigned by genotype
Gt(ROSA)26Sortm2Luo/Gt(ROSA)26Sortm2Luo
involves: 129S1/Sv * 129X1/SvJ
- reproductive system phenotype
- reduced female fertility (MGI Ref ID J:113338)
- homozygous females have low fertility
This mouse can be used to support research in many areas including:
GFP relatedCell Biology Research
Cell Cycle Regulation
Genes Regulating Growth and Proliferation
Research Tools
Cre-lox System (loxP-flanked Sequences: Test/Reporter)
Fluorescent Proteins
Genetics Research (Mutagenesis and Transgenesis: Cre-lox System)
Genetics Research (Tissue/Cell Markers: Cre-lox System)
Genetics Research (Tissue/Cell Markers: multiple)
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Gt(ROSA)26Sortm2Luo | ||
|---|---|---|---|
| Allele Name | targeted mutation 2, Liqun Luo | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | Gt(ROSA)26Sortm1(Dsred2/EGFP)Luo; MADM RG knockin; RG; | ||
| Mutation Made By | Liqun Luo, Stanford University, HHMI | ||
| Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ | ||
| ES Cell Line Name | R1 | ||
| ES Cell Line Strain | (129X1/SvJ x 129S1/Sv)F1-Kitl<+> | ||
| Site of Expression | when crossed to reciprocal strain (Stock No. 006041) and then to a cre strain, differential expression of the reporter protein(s) will be expressed in daughter cells, depending on chromatid segregation | ||
| 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 | RFP, Red Fluorescent Protein, jelly fish | ||
| Red Fluorescent Protein (RFP), derived from marine invertebrate organisms such as the soft coral Discosoma spp and reef coral, Heteractis crispa, is a versatile reporter molecule which has found use in many biological applications. The wild type protein, which is an obligate tetramer, is not well tolerated in mammalian systems. The original molecule has been modified in order to optimize expression to mammalian physiology (examples include monomeric RFP, mRFP1, DsRed, etc). | |||
| Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano | ||
| Chromosome | 6 | ||
| Gene Common Name(s) | AV258896; Gtrgeo26; Gtrosa26; R26; ROSA26; beta geo; expressed sequence AV258896; gene trap ROSA 26; gene trap ROSA b-geo 26; | ||
| Molecular Note | A construct containing the CMV beta-actin enhancer/promoter, SV40 poly(A) signal, and encoding the N-terminal part of Dsred2 (red fluorescent protein) and C-terminal part of EGFP which are separated by a beta-globin intron containing a loxP-flanked neomycin resistance gene, was knocked into ES cells at the ROSA26 locus. No protein is expressed from the chimeric locus in absence of Cre-mediated recombination because the coding sequences are interrupted by the intron in different reading frames. After recombination with the reciprocal mutation at the same locus carried by another mouse line, which restores the reading frames of EGFP and RFP, RNA splicing will remove the remaining single loxP sites, resulting in functional protein expression. [MGI Ref ID J:98961] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Gt(ROSA)26Sortm1Luo, tm2Luo, tm3Luo, tm4ACTB-tdTomato,-EGFP)Luo, STD PCR, vers. 1
Helpful Links
Optimizing PCR Protocols
References
References
Selected Reference(s)
Zong H; Espinosa JS; Su HH; Muzumdar MD; Luo L. 2005. Mosaic analysis with double markers in mice. Cell 121(3):479-92. [PubMed: 15882628] [MGI Ref ID J:98961]
Additional References
Gt(ROSA)26Sortm2Luo relatedLuo L. 2006. Gt(ROSA)26Sor<tm2Luo>- a marker for mosaic analysis MGI Direct Data Submission :. [MGI Ref ID J:113338]
Muzumdar MD; Luo L; Zong H. 2007. Modeling sporadic loss of heterozygosity in mice by using mosaic analysis with double markers (MADM). Proc Natl Acad Sci U S A 104(11):4495-500. [PubMed: 17360552] [MGI Ref ID J:120052]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, heterozygous mice are bred. Homozygous females produce less pups to weaning age compared to heterozygotes. Mating System Heterozygote x Heterozygote (Female x Male) Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
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Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $236.40 Female or Male Heterozygous for Gt(ROSA)26Sortm2Luo $291.90 Female or Male Homozygous for Gt(ROSA)26Sortm2Luo *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $472.80 Heterozygous for Gt(ROSA)26Sortm2Luo x Heterozygous for Gt(ROSA)26Sortm2Luo
Additional Supply Details
| Supply Notes |
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| Pricing for International shipping destinations |
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Weeks of Age Price* Gender Genotypes Provided Individual Mouse Price $307.40 Female or Male Heterozygous for Gt(ROSA)26Sortm2Luo $379.50 Female or Male Homozygous for Gt(ROSA)26Sortm2Luo *Price(s) in US dollars ($)
Pairs /Price* Pair Genotype $614.70 Heterozygous for Gt(ROSA)26Sortm2Luo x Heterozygous for Gt(ROSA)26Sortm2Luo
Additional Supply Details
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Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of ~nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within 48 hours of order placement. |
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Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 002448 129S1/SvImJ | (approximate) | |
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
General Terms and Conditions
See Terms of Use
The Jackson Laboratory's Genotype Promise
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 and Purchasing Information
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General Terms and Conditions
Effective September 26, 2007: License Requirements for Strains using Cre-lox Technology only apply in Canada, see Licenses for Strains using Cre-lox Technology.
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| phone: | 207-288-6470 |
| fax: | 207-288-6655 |
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