C57BL/6-Tg(Edn2-icre)9Jako/J

Stock number: 026975
Product name: edn2-iCre9
Research areas: Reproductive Biology Research, Research Tools

Description

These edn2-iCre transgenic mice have the mouse endothelin 2 (Edn2) promoter driving codon optimized iCre recombinase expression in the epidermis, ovary (capsule, stromal cells, corpora lutea, granulosa cells), testicular interstitium, coagulation gland (anterior prostate), and preputial gland.

Detailed description

The Edn2 gene encodes a member of the vasoconstrictive peptides and growth-promoting endothelin protein family, and which is predominantly expressed in the intestines, ovary, and uterus. These transgenic mice express iCre recombinase under the control of the mouse endothelin 2 promoter. When crossed with a strain containing loxP site flanked sequence of interest, Cre-mediated recombination results in deletion of the floxed sequence in the cre expressing tissues and in the ovary after gonadotropin stimulation. After treatment with human chorionic gonadotropin (hCG) to induce ovulation, Cre recombinase expression is detected in the ovary 11 and 12 hours after injection. iCre recombinase expression mimics the endogenous Edn2 gene expression pattern. Cre recombinase activity is also detected in developing dermis of the embryo starting at E15.5 and persisting through puberty. When bred to a reporter strain, the resulting mice exhibited strong Cre recombinase activity in the epidermis, ovary (capsule, stromal cells, corpora lutea, granulosa cells), testicular interstitium, coagulation gland (anterior prostate), preputial gland. Punctate Cre activity is detected in the oviduct epithelium, uterine smooth muscle, olfactory bulb, cerebral and cerebellar neurons, anterior and intermediate pituitary, cornea and retina of the eye, cardiac epicardium, scattered pneumocytes of the respiratory epithelium, salivary glands, liver parenchyma, pancreas, renal cortex corpuscles and tubules, adrenal, intestinal villi and tubular glands, and the detrusor muscle of the bladder. Mice that are homozygous for the transgene are viable and fertile.

Development

The mouse bacterial artificial chromosome (BAC) RP23-98J9, containing the entire Edn2 gene, was modified by insertion of sequence encoding icre, polyadenylation sequence, and a FRT site flanked NEO cassette, in front of the ATG start codon in exon 1. Arabinose-mediated FLP recombination (arabinose operon-controlled promoter FLP recombinase plasmid) treatment of the bacterial cells removed the FRT site flanked NEO cassette, resulting in one remaining FRT site. DNA from a correctly FLPe recombinase modified BAC clone was used as the source for the Edn2-iCre transgene, which was linearized and then microinjected into the pronucleus of C57BL/6 fertilized eggs. No other genes in the BAC were modified. Founder line 9 was subsequently established. The mice were maintained on a C57BL/6 background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Tg(Edn2-icre)9Jako
Name: transgene insertion 9, CheMyong Ko
MGI accession ID: MGI:5644453
Generation method: Transgenic
Attributes: Recombinase-expressing
Marker symbol: Tg(Edn2-icre)9Jako
Marker name: transgene insertion 9, CheMyong Ko
Chromosome: UN
Strain of origin: C57BL/6
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: When crossed with a strain containing loxP site flanked sequence of interest, Cre-mediated recombination results in deletion of the floxed sequence in the cre expressing tissues and in the ovary after gonadotropin stimulation.
Molecular note: The codon optimized icre and an FRT-flanked neomycin resistance cassette were inserted into exon 1 upstream of the translation initiation site of mouse Edn2. Flp-mediated recombination removed the selection cassette. Lines 9 and 12 were generated.