Arl13b-mCherry; Centrin2-GFP double transgenic mice express mCherry in cilia and EGFP in centrosomes making them useful when studying lineage specificity of primary cilia in the mouse embryo.
Kathryn V. Anderson, Memorial Sloan Kettering Cancer Center
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter) |
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Transgenic (Reporter) |
Arl13b-mCherry; Centrin2-GFP double transgenic mice have a murine ADP-ribosylation factor-like 13B (Arl13b) coding region of an Arl13b cDNA fused in-frame upstream of mCherry and driven by the pCAGGs promoter. They also express an enhanced green fluorescent protein-labeled human Centrin-2 (EGFP-CETN2). Double homozygous mice are viable and fertile. Colonists at The Jackson Laboratory have noted that double homozygous mice develop ear, eye, and nose sores by 12-14 weeks of age.
The Arl13b-mCherry fusion protein labels the membrane of primary cilia starting at E6.0 on epiblast cells, and by E8.0 labels all non-dividing epiblast-derived cells of the three germ layers, ectoderm, mesoderm and definitive endoderm. In contrast, Centrin 2-GFP labels centrioles in both inner cell mass (ICM) and trophectoderm cells in 32 cell blastocysts. At E10.5, mCherry-expressing cilia project from Centrin 2-GFP+ centrioles in cells throughout the embryo, including limb mesenchyme and neural epithelial cells. No ARL13B-mCherry+ cilia are associated with Centrin 2-GFP+ centrosomes in the cells of the extraembryonic-visceral-endoderm-derived layer of yolk sac cells.
The Arl13b-mCherry transgene was designed with a murine ADP-ribosylation factor-like 13B (Arl13b) cDNA fused in-frame upstream of a monomeric red fluorescent protein, mCherry, and driven by the pCAGGs promoter (which contains a chicken β-actin promoter and CMV immediate early enhancer). The transgene was microinjected into fertilized FVB oocytes and founder mice were bred to CB6-Tg(CAG-EGFP/CETN2)3-4Jgg/J transgenic mice that were purchased from The Jackson Laboratory as Stock No. 008234. These Centrin2-GFP mice express an enhanced green fluorescent protein-labeled human Centrin-2 (EGFP-CETN2) under the control of the chicken beta-actin promoter (with cytomegalovirus immediate early enhancer). Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.
Expressed Gene | CETN2, centrin 2, human |
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Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | GFP is expressed in a centrioles of the centrosome in every tissue examined from E14.5 through adult. |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Expressed Gene | Arl13b, ADP-ribosylation factor-like 13B, mouse, laboratory |
Site of Expression | Primary cilia on cells derived from the epiblast. |
Allele Name | transgene insertion 3-4, Joseph G Gleeson |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | Centrin 2-GFP; GFP-CETN2 |
Gene Symbol and Name | Tg(CAG-EGFP/CETN2)3-4Jgg, transgene insertion 3-4, Joseph G Gleeson |
Gene Synonym(s) | |
Promoter | ACTB, actin, beta, chicken |
Expressed Gene | CETN2, centrin 2, human |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | GFP is expressed in a centrioles of the centrosome in every tissue examined from E14.5 through adult. |
Strain of Origin | BALB/c x C57BL/6 |
Chromosome | UN |
Molecular Note | The GFP-CETN2 transgene was designed with a chicken beta-actin promoter (with cytomegalovirus immediate early enhancer and beta-actin intron) upstream of an EGFP-centrin 2 fusion protein (containing the full length human centrin 2 cDNA cloned in-frame into the C-terminal of enhanced green fluorescent protein), and followed by a beta-globin polyA sequence. Two lines were maintained (18-1 and 3-4) and line 3-4 was donated to The Jackson Laboratory. These lines show consistent, robust expression of GFP in centrosomes (particularly in cerebellar granule cells). |
Allele Name | transgene insertion 1, Kathryn Anderson |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | ARL13B-mCherry |
Gene Symbol and Name | Tg(CAG-Arl13b/mCherry)1Kvand, transgene insertion 1, Kathryn Anderson |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Expressed Gene | Arl13b, ADP-ribosylation factor-like 13B, mouse, laboratory |
Site of Expression | Primary cilia on cells derived from the epiblast. |
Strain of Origin | FVB/N |
Chromosome | UN |
Molecular Note | The transgenic construct is designed with a murine ADP-ribosylation factor-like 13B (Arl13b) cDNA fused in-frame upstream of a monomeric red fluorescent protein, mCherry, and driven by the pCAGGs promoter (which contains a chicken beta-actin promoter and CMV immediate early enhancer). Four founder lines were identified, only one exhibited consistent transgene expression. |
Double homozygous mice are viable and fertile. In 2019 colonists at The Jackson Laboratory noted that double homozygous mice develop ear, eye, and nose sores by 12-14 weeks of age. Therefore, our colony is maintained by breeding females homozygous for the ARL13B-mCherry transgene and hemizygous for the Centrin 2-GFP transgene (hom hemi) with males hemizygous for the ARL13B-mCherry transgene and homozygous for the Centrin 2-GFP transgene (hemi hom).
When using the Arl13b-mCherry; Centrin2-GFP mouse strain in a publication, please cite the originating article(s) and include JAX stock #027967 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous for Tg(CAG-Arl13b/mCherry)1Kvand, Hemizygous for Tg(CAG-EGFP/CETN2)3-4Jgg/, |
Frozen Mouse Embryo | STOCK Tg(CAG-Arl13b/mCherry)1Kvand Tg(CAG-EGFP/CETN2)3-4Jgg/ | $2595.00 |
Frozen Mouse Embryo | STOCK Tg(CAG-Arl13b/mCherry)1Kvand Tg(CAG-EGFP/CETN2)3-4Jgg/ | $2595.00 |
Frozen Mouse Embryo | STOCK Tg(CAG-Arl13b/mCherry)1Kvand Tg(CAG-EGFP/CETN2)3-4Jgg/ | $3373.50 |
Frozen Mouse Embryo | STOCK Tg(CAG-Arl13b/mCherry)1Kvand Tg(CAG-EGFP/CETN2)3-4Jgg/ | $3373.50 |
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