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| The MADM-11TG allele has the CMV enhancer/chicken beta-actin core promoter, the N-terminal portion of a red fluorescent protein (tdTomato), a beta-globin intronic sequence containing an frt site and a loxP-flanked neomycin resistance gene, and the C-terminal portion of a mutant enhanced green fluorescent protein (mut4-EGFP) all inserted into the Hipp11 locus on chromosome 11 (cytoband A1 at ~3cM between the Eif4enif1 and Drg1 loci). These MADM-11TG mutants are designed for MADM (mosaic analysis with double markers), and must be crossed to MADM-11GT mice harboring a reciprocal mutation at the same locus (see Stock No. 013749). This MADM system allows Cre- or FLP-mediated fluorescent labeling of daughter cells to ascertain lineal relationships and pleiotropic gene function in multicellular organisms. These mice may also be useful in studies of cell differentiation and mitosis. | |||||||||||||||||||
- Description
- Disease & phenotype
- Genes & alleles
- Genotyping
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- References
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Description
Strain Information
Former Names STOCK Tg(ACTB-tdTomato,-EGFP)11Luo/J (Changed: 01-FEB-13 ) Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System Homozygote x Homozygote (Female x Male) 07-NOV-11 Species laboratory mouse Generation F?+F4 (16-APR-13)
Generation DefinitionsDonating Investigator Dr. Liqun Luo, Stanford University, HHMI Description
Homozygous MADM-11TG mice are viable and fertile with no gross behavioral or observable abnormalities. The MADM-11TG allele has the CMV enhancer/chicken beta-actin core promoter, the N-terminal portion of a red fluorescent protein (tdTomato), a beta-globin intronic sequence containing an frt site and a loxP-flanked neomycin resistance gene, and the C-terminal portion of a mutant enhanced green fluorescent protein (mut4-EGFP) all inserted into the Hipp11 locus on chromosome 11 (cytoband A1 at ~3cM between the Eif4enif1 and Drg1 loci). These MADM-11TG mutants are designed for MADM (mosaic analysis with double markers), and must be crossed to MADM-11GT mice harboring a reciprocal mutation at the same locus (see Stock No. 013749). The resulting TG/GT offspring have one copy of each reciprocal mutation on homologous chromosomes ("trans-heterozygous"), and must also be bred to harbor a cre- or FLP-recombinase to induce fluorescent protein expression. Prior to Cre- or FLP-recombination, trans-heterozygous mutant mice do not have colored cells: the chimeric genes do not produce functional proteins because their coding sequences are interrupted by the beta-globin intron in different reading frames. After DNA replication (G2 phase) in double mutant mice, introduction of Cre- or FLP-recombinase 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 subsequent chromatid segregation (X or Z) determines daughter cell phenotype: recombinant sister chromatids into the same daughter cell (a G2-Z event) leads to double reporter expression or no reporter expression, while independent segregation into separate daughter cells (a G2-X event) leads to expression of either EGFP or tdTomato-MYC. If an additional targeted mutation of interest is introduced distal to the MADM-11 locus on chromosome 11, only homozygous cells will be singly labeled following G2 cre or FLP introduction. The homozygous mutant and wildtype cells can then be distinguished by which single reporter they express. Most heterozygous cells will be unlabeled, but some heterozygous cells will be yellow (both markers expressed). Reporter protein tissue specificity, expression levels, and frequency of recombination are thus determined by the promoter controlling Cre- or FLP-recombinase 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.Other important features of the MADM-11 system are listed below. Because of its placement ~3kb from the centromere, MADM-11 allows >99% of genes on chromosome 11 to be subjected to MADM-based mosaic analyses. Cre- or FLP-recombinase introduction in cell phase G0 or G1 results in double reporter expression. The donating investigator also reports the MADM-11 design has several advantages compared to the original MADM(-6) mice. Specifically, MADM-11 allows direct fluorescent visualization of both EGFP and tdTomato in live animals/cells: permitting genotypes of distinctly labeled cells in mosaic animals to be unequivocally determined prior to fixation and/or immunostaining. Also, MADM-11 contains both loxP sites and a frt site; allowing the induction of MADM-labeling by either Cre recombinase or FLP recombinase. In addition, the interchromosomal recombination rate in MADM-11 is markedly increased compared with the original MADM(-6) system; allowing greater temporal control of clone induction if using concomitantly with an inducible Cre recombinase (or Flp recombinase).
Development
The TG MADM targeting construct was designed with a CMV enhancer/chicken beta-actin core promoter (pCA), the N-terminal portion of a red fluorescent protein (tdTomato; aa 1-3), a beta-globin intronic sequence containing an frt site and loxP-flanked neomycin resistance gene, the C-terminal portion of a mutant enhanced green fluorescent protein (mut4-EGFP; aa 274-724), and an SV40 T-antigen poly(A) signal. This entire TG MADM construct was inserted into the Hipp11 locus on chromosome 11 (cytoband A1 at ~3cM between the Eif4enif1 and Drg1 loci) via electroporation of (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were microinjected into recipient blastocysts and chimeric progeny were bred with C57BL/6 mice to establish the MADM-11TG colony. MADM-11TG mice were bred with CD1 outbred mice for at least three generations. MADM-11TG mice on a mixed CD1;C57BL/6;129 genetic background were sent to The Jackson Laboratory Repository. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (358 strains)
017530 STOCK Iis2tm2(ACTB-tdTomato,-EGFP)Luo Trp53tm1Tyj Nf1tm1Par/J
010939 B6.Cg-Tg(ACTB-UPF1*R844C)581Hcd/J 005703 B6.Cg-Tg(ACTFLPe)9205Dym/J 009686 B6.Cg-Tg(Actb-TNFRSF6B)754Jwu/J 008226 B6.FVB-Tg(CAG-EGFP,-ALPP)2.6Ggc/J 003800 B6;SJL-Tg(ACTFLPe)9205Dym/J 005145 C57BL/6-Tg(CAG-OVA)916Jen/J 005863 C57BL/6J-Tg(ACTB-DDAH1)1Jpck/J 002981 DBA/2-Tg(xstpx-lacZ)36And/J 003376 FVB/N-Tg(ACTB-cre)2Mrt/J 008200 FVB/N-Tg(CAG-EGFP,-ALPP)2.6Ggc/J 013749 STOCK Iis2tm1(ACTB-EGFP,-tdTomato)Luo/J 005438 STOCK Tg(CAG-Bgeo,-DsRed*MST)1Nagy/J
013749 STOCK Iis2tm1(ACTB-EGFP,-tdTomato)Luo/J
Additional Web Information
FLP-FRT Systems
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Phenotype
Phenotype Information
This mouse can be used to support research in many areas including:
GFP relatedCell Biology Research
Cell Cycle Regulation
Genes Regulating Growth and Proliferation
Neurobiology Research
Cre-lox System
loxP-flanked Sequences
loxP-flanked Sequences: Test/Reporter
Fluorescent protein expression in neural tissue
Research Tools
Cardiovascular Research
Cre-lox System
Cell Biology Research
Cre-lox System
loxP-flanked Sequences
loxP-flanked Sequences: Test/Reporter
Developmental Biology Research
Cre-lox System
Diabetes and Obesity Research
loxP
FLP-FRT System
FRT-flanked Sequences
FRT-flanked Sequences: Test/Reporter
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers
Tissue/Cell Markers: Cre-lox System
Neurobiology Research
cell marker
Reproductive Biology Research
Cre-lox System
Research Tools
Fluorescent Proteins
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Iis2tm2(ACTB-tdTomato,-EGFP)Luo | ||
|---|---|---|---|
| Allele Name | targeted mutation 2, Liqun Luo | ||
| Allele Type | Targeted (Reporter) | ||
| Common Name(s) | MADM-11-TG; MADM-11TG; TG: tdTN-termGFPC-term; Tg(ACTB-tdTomato,-EGFP)11Luo; | ||
| Mutation Made By | Dr. Liqun Luo, Stanford University, HHMI | ||
| Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl<+> | ||
| Site of Expression | Mutant daughter cells express GFP, while the sibling wildtype cells are labeled with RFP. | ||
| 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, coral | ||
| 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). | |||
| Promoter | ACTB, actin, beta, human | ||
| Molecular Note | The TG MADM targeting construct was designed with a CMV enhancer/chicken beta-actin core promoter (pCA), the N-terminal portion of a red fluorescent protein (tdTomato; aa 1-3), a beta-globin intronic sequence containing an frt site and loxP-flanked neomycin resistance gene, the C-terminal portion of a mutant enhanced green fluorescent protein (mut4-EGFP; aa 274-724), and an SV40 T-antigen poly(A) signal. This entire TG MADM construct was inserted into the "Hipp11" locus on chromosome 11 (cytoband A1 at ~3cM between the Eif4enif1 and Drg1 loci). [MGI Ref ID J:166047] | ||
| Gene Symbol and Name | Iis2, intergenic insertion site 2 | ||
| Chromosome | 11 | ||
| Gene Common Name(s) | H11; Hipp11; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Generic GFP/EGFP qPCR, QPCR
Tg(ACTB-tdTomato,-EGFP)11Luo, Melt Curve Analysis
Tg(ACTB-tdTomato,-EGFP)11Luo, Standard PCR
Fluorescent Proteins -- Generic GFP, QPCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
Selected Reference(s)
Hippenmeyer S; Youn YH; Moon HM; Miyamichi K; Zong H; Wynshaw-Boris A; Luo L. 2010. Genetic mosaic dissection of lis1 and ndel1 in neuronal migration. Neuron 68(4):695-709. [PubMed: 21092859] [MGI Ref ID J:166047]
Additional References
Iis2tm2(ACTB-tdTomato,-EGFP)Luo relatedLiang H; Xiao G; Yin H; Hippenmeyer S; Horowitz JM; Ghashghaei HT. 2013. Neural development is dependent on the function of specificity protein 2 in cell cycle progression. Development 140(3):552-61. [PubMed: 23293287] [MGI Ref ID J:194076]
Liu C; Sage JC; Miller MR; Verhaak RG; Hippenmeyer S; Vogel H; Foreman O; Bronson RT; Nishiyama A; Luo L; Zong H. 2011. Mosaic analysis with double markers reveals tumor cell of origin in glioma. Cell 146(2):209-21. [PubMed: 21737130] [MGI Ref ID J:174616]
Liu S; Liu Y; Hao W; Wolf L; Kiliaan AJ; Penke B; Rube CE; Walter J; Heneka MT; Hartmann T; Menger MD; Fassbender K. 2012. TLR2 is a primary receptor for Alzheimer's amyloid beta peptide to trigger neuroinflammatory activation. J Immunol 188(3):1098-107. [PubMed: 22198949] [MGI Ref ID J:180762]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, homozygous mice may be bred together. Mating System Homozygote x Homozygote (Female x Male) 07-NOV-11 Diet Information LabDiet® 5K52/5K67
Pricing and Purchasing
Pricing, Supply Level & Notes, Controls
| Pricing for USA, Canada and Mexico shipping destinations |
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Live Mice
Price per mouse (US dollars $) Gender Genotypes Provided Individual Mouse $232.00 Female or Male Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo
Price per Pair (US dollars $) Pair Genotype $464.00 Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo x Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo 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 Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo
Price per Pair (US dollars $) Pair Genotype $603.20 Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo x Homozygous for Iis2tm2(ACTB-tdTomato,-EGFP)Luo 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.
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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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