C57BL/6N-Tg(Slc6a3-EGFP/Rpl10a)I17-11Pggd/J

Stock number: 012365
Product name: Dat bacTRAP
Research areas: Neurobiology Research, Research Tools

Description

These Dat bacTRAP mice express a EGFP/Rpl10a, green fluorescent protein-tagged ribosomal protein L10a, transgene under the control of the mouse Slc6a3 gene, in midbrain dopaminergic neurons. These mice may be useful in translational profiling (TRAP, translating ribosome affinity purification) studies related to Parkinson's Disease.

Detailed description

These Dat bacTRAP transgenic mice express Enhanced Green Fluorescent Protein tagged ribosomal protein L10A under the control of the mouse Slc6a3 gene promoter. The Slc6a3 gene, also known as Dat, encodes a dopamine transporter. Transgene expression is detected in midbrain dopaminergic neurons. Hemizygous mice of founder line I17-11 carry 6 to 10 copies of the transgene, in tandem, on chromosome 9q, as revealed by fluorescent in situ hybridization (FISH) analysis. This strain serves as a translating ribosome affinity purification (TRAP) strain for isolating translated mRNAs from midbrain dopaminergic neurons for use in translational profiling. Hemizygotes are viable and fertile. Homozygous viability/fertility has not been tested (May 2017).

Development

The mouse BAC (bacterial artificial chromosome) RP24-269I17 containing the entire Slc6a3 gene, was modified by inserting cDNA encoding EGFP/Rpl10a (Enhanced Green Fluorescent Protein fused to ribosomal protein L10A) and a 3’ polyA sequence upstream of the endogenous Slc6a3 ATG initiating codon. DNA from a modified BAC clone was used as the source for the Dat bacTRAP transgene, which was microinjected into the pronucleus of C57BL/6NCrl fertilized eggs. The resulting transgenic mice from founder line I17-11 were subsequently backcrossed to C57BL/6NCrl for 10 generations prior to arrival at The Jackson Laboratory. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6NCrl genetic background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (Stock No. 005304).

Allele

Symbol: Tg(Slc6a3-EGFP/Rpl10a)I17-11Pggd
Name: transgene insertion I17-11, Paul Greengard
MGI accession ID: MGI:5897802
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Slc6a3-EGFP/Rpl10a)I17-11Pggd
Marker name: transgene insertion I17-11, Paul Greengard
Chromosome: UN
Strain of origin: C57BL/6NCrl
Expressed genes: Rpl10a,ribosomal protein L10A,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: EGFP labeling is observed in dopaminergic cell bodies.
Molecular note: The mouse BAC (bacterial artificial chromosome) RP24-269I17 containing the entire Slc6a3 gene, was modified by inserting cDNA encoding EGFP/Rpl10a (Enhanced Green Fluorescent Protein fused to ribosomal protein L10A) and a 3' polyA sequence upstream of the endogenous Slc6a3 ATG initiating codon. EGFP expressionis detected in dopaminergic cell bodies and does not extend to the projections of these neurons.