Ple137-EGFP (Negative)

Ple137-EGFP Description:

The Ple137 promoter construct is derived from the promoter region of the Orphan nuclear receptor NR2E1 (NR2E1) gene. The Pleiades Promoter Project constructed a 3587 bp MiniPromoter from two downstream putative regulatory elements and a subsection of the NR2E1 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 pEMS1353 with Ple137 driving EGFP. The Ple137-EGFP (pEMS1353) 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 (mEMS1675, mEMS1676, mEMS1808, and mEMS1809) were PCR validated, and chimeras were generated using mEMS1808.

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

Ple137-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).

Ple137 MiniPromoter Resources:

Ple137-EGFP
	plasmid pEMS1353 (in backbone pEMS1306)
	ESCs mEMS1675, mEMS1676, mEMS1808, mEMS1809
	mice generated using ESC mEMS1808
Resource Files
	Ple137 pEMS1353 Sequence	Jones lab
	Ple137 pEMS1353 MiniPromoter Design	Wasserman lab
	Ple137 pEMS1353 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple137 pEMS1353 Vector Map	Holt lab
	Ple137 pEMS1353 Genotyping Assay	Simpson lab

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