Ple221-EGFP (Negative)

Ple221-EGFP Description:

The Ple221 promoter construct is derived from the promoter region of the Tachykinin 2 (Tac3) gene. The Pleiades Promoter Project created a 1305 bp MiniPromoter derived from a candidate intronic regulatory region along with 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 pEMS1184 with Ple221 driving EGFP. The Ple221-EGFP (pEMS1184) 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) mEMS1204.03B. Four ESC lines (mEMS1485, mEMS1491, mEMS1492, and mEMS1493) were PCR validated. Chimeras were generated using mEMS1492.

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

Ple221-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).

Ple221 MiniPromoter Resources:

Ple221-EGFP
	plasmid pEMS1184 (in backbone pEMS1306)
	ESCs mEMS1485, mEMS1491, mEMS1492, mEMS1493
	mice generated using ESC mEMS1492
Resource Files
	Ple221 pEMS1184 Sequence	Jones lab
	Ple221 pEMS1184 MiniPromoter Design	Wasserman lab
	Ple221 pEMS1184 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple221 pEMS1184 Vector Map	Holt lab
	Ple221 pEMS1184 Genotyping Assay	Simpson lab
	Ple221 pEMS1184 Genotyping Assay  (Requires VectorNTI Sofware)	Holt lab

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