Ple220-EGFP (Negative)

Ple220-EGFP Description:

The Ple220 promoter construct is derived from the promoter region of the tachykinin 2 (TAC3) gene. The Pleiades Promoter Project created a 990 bp MiniPromoter derived from a downstream candidate regulatory region of the TAC3 gene, along with a subsection of the TAC3 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 pEMS1183 with Ple220 driving EGFP. The Ple220-EGFP (pEMS1365) 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) mEMS1204.03A. Four ESC lines (mEMS1488, mEMS1489, mEMS1490, and mEMS1501) were PCR validated. Chimeras were generated using mEMS1488.

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

Ple220-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).

Ple220 MiniPromoter Resources:

Ple220-EGFP
	plasmid pEMS1183 (in backbone pEMS1306)
	ESCs mEMS1488, mEMS1489, mEMS1490, mEMS1501
	mice generated using ESC mEMS1488
Resource Files
	Ple220 pEMS1183 Sequence	Jones lab
	Ple220 pEMS1183 MiniPromoter Design	Wasserman lab
	Ple220 pEMS1183 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple220 pEMS1183 Vector Map	Holt lab
	Ple220 pEMS1183 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

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