FVB/N-Tg(tetO-Ube3a*2)884Svd/J

Stock number: 026279
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools

Description

These TRE-mUbe3a_v2[884] transgenic mice allow Tet-Off/Tet-On expression of a FLAG-tagged Ube3a splice variant 2 protein, in addition to normal expression of endogenous Ube3a. These mice are useful to study temporally-controlled, tissue-specific Ube3a variant 2 overexpression as a model for maternal 15q11-13 duplication (dup15) and triplication (isodicentric extranumerary chromosome, idic15); both are associated with human autism spectrum disorder.

Detailed description

The TRE-Ube3av2 transgene has a modified Tet response element (TRE or tetO) upstream of a cDNA sequence encoding a functional, full-length mouse ubiquitin protein ligase E3A transcript variant 2 protein with N-terminal FLAG tag (FLAG-Ube3a_v2). Transgenic mice from founder line 884 (TRE-mUbe3a_v2[884]) are described here.


When these TRE-mUbe3a_v2[884] transgenic mice are bred with another mouse expressing tetracycline-controlled transactivator protein (tTA) or reverse tetracycline-controlled transactivator protein (rtTA), FLAG-Ube3a_v2 expression in the resulting double mutant offspring can be regulated with tetracycline or its analog doxycycline (dox). As designed, TRE-mUbe3a_v2[884] transgenic mice have no reported levels of FLAG-Ube3a_v2 expression in absence of tTA. When TRE-mUbe3a_v2[884] transgenic mice are bred to Camk2a-tTA transgenic mice (see Stock Nos. 003010 / 007004), the resulting CaMKIIα-mUbe3a_v2[884] double transgenic mice have FLAG-Ube3a_v2 expression in forebrain neurons.


TRE-mUbe3a_v2[884] transgenic mice have normal expression of endogenous Ube3a. Importantly, the endogenous Ube3a gene is only expressed from the maternal chromosome in neurons; a result of paternal imprinting (the antisense transcript functions to silence paternal expression in brain). Because the TRE-Ube3av2 transgene lacks the transcription initiation site of the antisense transcript, expression of FLAG-Ube3a_v2 is independent of parent-of-origin or sex of the animal.


Hemizygous TRE-mUbe3a_v2[884] transgenic mice are viable and fertile. The donating investigator has not tried to generate homozygous mice to date (December 2014).

Development

The TRE-Ube3av2 transgene was designed in the laboratory of Dr. Scott V. Dindot (Texas A&M University). This transgene has (from 5' to 3') a modified Tet response element (TRE or tetO; described below), a mouse ubiquitin protein ligase E3A (Ube3a) cDNA sequence encoding transcript variant 2 (NM_011668.2), and an SV40 polyA signal. The Ube3a sequence has a FLAG epitope tag attached to its 5' end. The ~3.5 kbp TRE-Ube3av2 transgene was microinjected into FVB/N embryos. Transgenic founder animals were identified by genotyping, and then bred to FVB/NJ. TRE-mUbe3a_v2 founder mice were subsequently bred to FVB/NJ inbred mice. Mice from founder line 884 (TRE-mUbe3a_v2[884]) were propagated and maintained by breeding to FVB/NJ inbred mice and/or wildtype (noncarrier) siblings for several generations. Albino males were sent to The Jackson Laboratory Repository in 2014. Upon arrival, sperm was cryopreserved. To establish our living colony, an aliquot of the frozen sperm was used to fertilize oocytes from FVB/NJ inbred females (Stock No. 001800).

The TRE promoter used here is Tet-responsive P_tight; containing a modified Tet response element (TRE_mod) composed of seven direct 36 bp repeats (each containing the 19-bp tet operator sequence [tetO]). TRE_mod is just upstream of a minimal human cytomegalovirus promoter (P_minCMVΔ), which lacks the enhancer that is part of the complete CMV promoter. Consequently, P_tight is silent in the absence of binding of TetR or rTetR to the tetO sequences.

Allele

Symbol: Tg(tetO-Ube3a*2)884Svd
Name: transgene insertion 884, Scott Dindot
MGI accession ID: MGI:5606057
Generation method: Transgenic
Attributes: Inducible; Inserted expressed sequence
Marker symbol: Tg(tetO-Ube3a*2)884Svd
Marker name: transgene insertion 884, Scott Dindot
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: Ube3a,ubiquitin protein ligase E3A,mouse, laboratory
Site of expression: Expression of FLAG-tagged Ube3a can be regulated with tetracycline or its analog doxycycline when combined with a mutant carrying tTA or rtTA.
Molecular note: This transgene has (from 5' to 3') a modified Tet response element (TRE or tetO), a mouse ubiquitin protein ligase E3A (Ube3a) cDNA sequence encoding transcript variant 2 (NM_011668.2), and an SV40 polyA signal. The Ube3a sequence has a FLAG epitope tag attached to its 5' end.