B6;FVB-Tg(Hcrt-EGFP/Rpl10a)JD218Jdd/J

Stock number: 028619
Product name: Hcrt bacTRAP
Research areas: Neurobiology Research, Research Tools

Description

This bacTRAP line expresses EGFP-RPL10a from the mouse Hcrt promoter. Ribosome-bound mRNAs from Hcrt-expressing neurons of the hypothalamus can be purified using EGFP as an affinity tag.

Detailed description

Hypocretin (orexin; Hcrt)-containing neurons of the hypothalamus are essential for the normal regulation of sleep and wake behaviors and have been implicated in feeding, anxiety, depression, and reward.

This bacTRAP line expresses EGFP-RPL10a from the mouse Hcrt promoter. Ribosome-bound mRNAs from Hcrt-expressing neurons of the hypothalamus can be purified using EGFP as an affinity tag. GFP expression, as determined by confocal immunofluorescent microscopy, is reported in greater than 85% of neurons with detectable Hcrt expression and there are no instances of EGFP-RPL10a expression in Hcrt-negative cells. No EGFP-RPL10a expression is reported elsewhere in the adult brain.

Development

EGFP-RPL10a was inserted at the translation start site of the Hcrt gene encoded by BAC RP23-258L05. The modified BAC was injected into FVB eggs. Resultant founder mice (founder line JD218) were crossed with C57BL/6J animals for 3 or 4 generations by the donating lab.

Allele

Symbol: Tg(Hcrt-EGFP/Rpl10a)JD218Jdd
Name: transgene insertion JD218, Joseph D Dougherty
MGI accession ID: MGI:5496432
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Hcrt-EGFP/Rpl10a)JD218Jdd
Marker name: transgene insertion JD218, Joseph D Dougherty
Chromosome: UN
Strain of origin: FVB
Expressed genes: EGFP/Rpl10,enhanced green fluorescent protein; ribosomal protein 10,
Site of expression: EGFP/Rpl10a fusion protein is expressed in neurons with detectable Hcrt expression.
Molecular note: BAC RP23-258L05, was modified by insertion of an EGFP/RPL10a fusion construct at the translation start site of the Hcrt gene.