Ple46-EGFP (Negative)

Ple46-EGFP Description:

The Ple46 promoter construct is derived from the promoter region of the CX3 chemokine receptor 1 (CX3CR1) gene. The Pleiades Promoter Project constructed a 3964 bp MiniPromoter from three candidate regulatory regions of the CX3CR1 gene along with a subsection of the CX3CR1 promoter and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1307 (EGFP reporter flanked by full frt sites) to make plasmid pEMS1151 with Ple46 driving EGFP. The Ple46-EGFP (pEMS1151) 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 (mEMS3086, mEMS3099, mEMS3108, and mEMS3109) were PCR validated. Chimeras were generated using mEMS3109.

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

Ple46-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).

Ple46 MiniPromoter Resources:

Ple46-EGFP
	plasmid pEMS1151 (in backbone pEMS1307)
	ESCs mEMS3086, mEMS3099, mEMS3108, mEMS3109
	mice generated using ESC mEMS3109
Resource Files
	Ple46 pEMS1151 Sequence	Jones lab
	Ple46 pEMS1151 MiniPromoter Design	Wasserman lab
	Ple46 pEMS1151 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple46 pEMS1151 Vector Map	Holt lab

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