STOCK Tg(CAG-Arl13b/mCherry)1Kvand Tg(CAG-EGFP/CETN2)3-4Jgg/KvandJ

Stock number: 027967
Product name: Arl13b-mCherry; Centrin2-GFP
Research areas: Research Tools

Description

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.

Detailed description

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.

Development

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.

Allele

Symbol: Tg(CAG-EGFP/CETN2)3-4Jgg
Name: transgene insertion 3-4, Joseph G Gleeson
MGI accession ID: MGI:3793421
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(CAG-EGFP/CETN2)3-4Jgg
Marker name: transgene insertion 3-4, Joseph G Gleeson
Chromosome: UN
Strain of origin: BALB/c x C57BL/6
Expressed genes: CETN2,centrin 2,human; 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.
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

Symbol: Tg(CAG-Arl13b/mCherry)1Kvand
Name: transgene insertion 1, Kathryn Anderson
MGI accession ID: MGI:5645691
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(CAG-Arl13b/mCherry)1Kvand
Marker name: transgene insertion 1, Kathryn Anderson
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: RFP,Red Fluorescent Protein,coral; Arl13b,ADP-ribosylation factor-like 13B,mouse, laboratory
Site of expression: Primary cilia on cells derived from the epiblast.
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.