Ple219-EGFP (Negative)

Ple219-EGFP Description:

The Ple219 promoter construct is derived from the promoter region of the Tachykinin 2 (TAC3) gene. The Pleiades Promoter Project constructed a 1802 bp MiniPromoter from three upstream putative regulatory elements and a subsection of the TAC3 promoter and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1182 with Ple219 driving EGFP. The Ple219-EGFP (pEMS1182) was introduced by homologous recombination as a single copy insertion into the Hprt1b-m3 locus in the X chromosome of mouse embryonic stem cells (ESCs) mEMS1204. Four ESC lines (mEMS1486, mEMS1487, mEMS1505, and mEMS1528) were PCR validated, and chimeras were generated using mEMS1487.

Note: Promoterless negative controls (pEMS1302, pEMS1306, pEMS1313) have been generated, as well as CAG positive controls (pEMS1157, pEMS1277 & pEMS1488).

Ple219-EGFP Expression Pattern:

No EGFP expression was detected above background in native fluorescence under epi or confocal microscopy. Anti GFP immunostaining (rabbit polyclonal, Invitrogen) did not detect specific GFP expression above background across numerous brain regions (brightfield).

Ple219 MiniPromoter Resources:

Ple219-EGFP
	plasmid pEMS1182 (in backbone pEMS1306)
	ESCs mEMS1486, mEMS1487, mEMS1505, mEMS1528
	mice generated using ESC mEMS1487
Resource Files
	Ple219 pEMS1182 MiniPromoter Design	Wasserman lab
	Ple219 pEMS1182 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple219 pEMS1182 Vector Map	Holt lab
	Ple219 pEMS1182 Genotyping Assay	Simpson lab

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