CAG-RFP-EGFP-LC3 transgenic mice express RFP and EGFP in a pH-dependent manner in phagocytic cells following ischemic injury. They are useful for studying the initiation, progression, and resolution of autophagy in a variety of tissues following ischemic injury.
Joseph Hill, The University of Texas Southwestern Medical Center
Genetic Background | Generation |
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?+pN3
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Allele Type |
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Transgenic (Reporter, Inserted expressed sequence) |
Starting at:
$255.00 Domestic price for female 4-week |
333.51 Domestic price for breeder pair |
CAG-RFP-EGFP-LC3 transgenic mice have CAG promoter/enhancer sequences driving expression of a red fluorescent protein (RFP), an enhanced green fluorescent protein (EGFP), and a microtubule-associated protein 1 light chain 3 alpha (Map1lc3a or LC3) gene. LC3 serves as a marker for phagosomes in a wide variety of tissues. The dual expression of two fluorophores allows tracking of different types of phagocytic cellular compartments depending on the acidity of the environment. RFP is stable in acidic pH (pKa 4.5) while EGFP (pKa 5.9) is quenched in the acidic lysosomal environment, allowing autophagosomes to be distinguished from autolysosomes. Combined GFP and RFP fluorescence yields a yellow signal within high pH phagophores and autophagosomes, while EGFP is quenched in autolysosomes, and they emit only an RFP signal. Following ischemia-reperfusion injury, EGFP expression reaches a peak at 24 hours and returns to baseline levels at 3 days. Meanwhile, RFP expression also peaks at 24 hours but persists at high levels for 3 days before returning to baseline levels by 7 days. CAG-RFP-EGFP-LC3 mice are useful for studying the progression and recovery from ischemic injury in a variety of tissues. Hemizygotes are viable and fertile. The donating investigator did not try to make a homozygous colony.
The CAG-RFP-EGFP-LC3 transgene was designed with a human cytomegalovirus immediate early promoter enhancer with chicken beta-actin/rabbit beta-globin hybrid promoter (CAG; from the pCAGGS vector) driving expression of a red fluorescent protein (RFP), an enhanced green fluorescent protein (EGFP), and a microtubule-associated protein 1 light chain 3 alpha (Map1lc3a or LC3) gene. The transgene was microinjected into fertilized C57BL/6 oocytes and the donating investigator reported that founder mice were bred to C57BL/6 mice (see SNP note below). Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. One of the 27 markers throughout the genome was segregating from an unknown source. All 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Expressed Gene | RFP, Red Fluorescent Protein, coral |
Expressed Gene | Map1lc3b, microtubule-associated protein 1 light chain 3 beta, mouse, laboratory |
Site of Expression | Expression occurs in a variety of tissues. |
Allele Name | transgene insertion 1, Joseph Hill |
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Allele Type | Transgenic (Reporter, Inserted expressed sequence) |
Allele Synonym(s) | CAG-RFP-EGFP-LC3 |
Gene Symbol and Name | Tg(CAG-RFP/EGFP/Map1lc3b)1Hill, transgene insertion 1, Joseph Hill |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | GFP, Green Fluorescent Protein, |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Expressed Gene | Map1lc3b, microtubule-associated protein 1 light chain 3 beta, mouse, laboratory |
Site of Expression | Expression occurs in a variety of tissues. |
Strain of Origin | C57BL/6 |
Chromosome | UN |
Molecular Note | The transgene was designed with a human cytomegalovirus immediate early promoter enhancer with chicken beta-actin/rabbit beta-globin hybrid promoter (CAG; from the pCAGGS vector) driving expression of a fusion of a red fluorescent protein (RFP) with a Strep-tag sequence to facilitate isolation, an enhanced green fluorescent protein (EGFP), and a microtubule-associated protein 1 light chain 3 beta (Map1lc3b or LC3) gene. |
When maintaining a live colony, hemizygous mice may be bred with wildtype (non-carrier) mice from the colony. The donating investigator did not try to make the colony homozygous.
When using the CAG-RFP-EGFP-LC3 mouse strain in a publication, please cite the originating article(s) and include JAX stock #027139 in your Materials and Methods section.
Service/Product | Description | Price |
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Hemizygous or Non carrier for Tg(CAG-RFP/GFP/Map1lc3b)1Hill |
Frozen Mouse Embryo | C57BL/6-Tg(CAG-RFP/EGFP/Map1lc3b)1Hill/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Tg(CAG-RFP/EGFP/Map1lc3b)1Hill/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | C57BL/6-Tg(CAG-RFP/EGFP/Map1lc3b)1Hill/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | C57BL/6-Tg(CAG-RFP/EGFP/Map1lc3b)1Hill/J Frozen Embryo | $3373.50 |
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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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