Ple94-EGFP (Negative)

Ple94-EGFP Description:

The Ple94 promoter construct is derived from the promoter region of the G-protein coupled receptor 88 (GPR88) gene. The Pleiades Promoter Project created a 3053 bp MiniPromoter derived from two upstream candidate regulatory regions, along with a segment of the promoter and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1102 with Ple94 driving EGFP. The Ple94-EGFP (pEMS1102) 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 (mEMS1549, mEMS1550, mEMS1551, and mEMS1562) were PCR validated. Chimeras were generated using mEMS1550.

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

Ple94-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).

Ple94 MiniPromoter Resources:

Ple94-EGFP
	plasmid pEMS1102 (in backbone pEMS1306)
	ESCs mEMS1549, mEMS1550, mEMS1551, mEMS1562
	mice generated using ESC mEMS1550
Resource Files
	Ple94 pEMS1162 Sequence	Jones lab
	Ple94 pEMS1162 MiniPromoter Design	Wasserman lab
	Ple94 pEMS1162 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple94 pEMS1162 Vector Map	Holt lab
	Ple94 pEMS1162 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

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