C57BL/6J-Tg(Adamts18-rtTA)1Amc/J

Stock number: 034205
Product name: Adamts18en-rtTA (AE-rtTA)
Research areas: Research Tools

Description

AE-rtTA mice are a Tet-On tool that allows conditional, doxycycline-inducible expression of the reverse tetracycline-controlled transactivator (rtTA) protein in the kidney ureteric bud tip using the Adamts18 enhancer and a minimal promoter.

Detailed description

Adamts18en-rtTA (AE-rtTA) transgenic mice allow Tet-inducible manipulation specifically within the kidney ureteric bud (UB) tip population during development. The transgene utilizes an Adamts18 enhancer discovered to be active in the kidney but inactive in the lung. Adamts18 (a disintegrin-like and metallopeptidase (reprolysin type) with thrombospondin type 1 motif, 18) encodes a secreted protease. This ureteric bud tip (UBT) enhancer drives the transcription of rtTA (reverse tetracycline-controlled transactivator). Homozygous mice are viable and fertile. When mated to a second transgenic strain carrying a gene of interest under the regulatory control of a tetracycline-responsive promoter element (TRE or tetO), expression of the target gene in mature adipocytes is induced with administration of the tetracycline analog, doxycycline (dox).

For example, when combined with STOCK Tg(tetO-HIST1H2BJ/GFP)47Efu/J (Stock No. 005104), embryos from doxycycline treated females can be used for live imaging of embryonic kidney explants and the observation of ureteric bud branching events at the cellular level.

Furthermore, crossing AE-rtTA mice to B6.Cg-Tg(tetO-DTA)1Gfi/J (Stock No. 008468) and treating pregnant females with doxycycline, results in embryonic tip ablation through the production of Diphtheria Toxin A (DTA) at the onset of ureteric bud outgrowth, which affects the subsequent development of the kidney, resulting in kidney agenesis, duplex kidneys, and double ureters.

Development

The transgene has (from 5' to 3'), an insulator sequence, Adamts18 intronic enhancer region from between exons 2 and 3 (Chr8:113,848,193-113,845,196, mouse assembly mm10), an Hsp68 minimal promoter sequence, the rtTA gene and a polyA sequence. The linearized construct was introduced by pronuclear injection into blastocysts from C57BL/6J mice. Founder mice was mated to C57BL/6J and maintained on the C57BL/J background and maintained by hemizygous intercrosses. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Tg(Adamts18-rtTA)1Amc
Name: transgene insertion 1, Andrew P McMahon
MGI accession ID: MGI:6376232
Generation method: Transgenic
Attributes: Transactivator
Synonyms: AE-rtTA
Marker symbol: Tg(Adamts18-rtTA)1Amc
Marker name: transgene insertion 1, Andrew P McMahon
Chromosome: UN
Strain of origin: C57BL/6J
Site of expression: cells of the kidney ureteric bud

Molecular note: The transgene has (from 5' to 3'), an insulator sequence, the Adamts18 intronic enhancer region from between exons 2 and 3 (Chr8:113,848,193-113,845,196, mouse assembly mm10), an Hsp68 minimal promoter sequence, the rtTA gene and a polyA sequence.