Ple167-EGFP/cre (Positive)

Ple167-EGFP/cre Description:

The Ple167 promoter construct is derived from the promoter region of the Pogo Transposable Element With ZNF Domain (POGZ) gene. The Pleiades Promoter Project created a 1200 bp MiniPromoter derived from isolated a segment of the basal promoter element and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1302 (EGFP/cre reporter flanked by minimal frt sites) to make plasmid pEMS1091 with Ple167 driving EGFP/cre. The Ple167-EGFP/cre (pEMS1091) 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) mEMS21TG2A. Four ESC lines (mEMS670, mEMS672, mEMS681, and mEMS688) were PCR validated. Chimeras were generated using mEMS681.

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

Ple167-EGFP/cre Expression Pattern:

Expression of the Ple167 MiniPromoter driving cre causes rearrangement of the Gt(ROSA)26Sortm1Sor reporter locus, thus producing consistently strong ßGal activity widely throughout the brain with relatively little regional specificity. Cells are present in the cortex, striatum, hippocampus, thalamus, midbrain, brainstem, and in the cerebellum. It is striking that the lacZ-positive cells represent only a small percentage of the cells in the brain with a seemingly unrestricted pattern of expression with respect to cell type. It appears that whole cortical columns are labeled, as well as subpopulations of cells in all hippocampal areas and even subpopulations of cerebellar granule cells and Purkinje cells are labeled indicating no apparent organizing principles in the expression of the MiniPromoter. EGFP expression, however, was not detected in these brains by either native fluorescence or immunocytochemistry. The absence of EGFP expression and the small number of total cells labeled suggests the historical nature of the functional activation of the promoter at an earlier time in development.

Ple167 MiniPromoter Resources:

Ple167-EGFP/cre
	plasmid pEMS1091 (in backbone pEMS1302)
	ESCs mEMS670, mEMS672, mEMS681, mEMS688
	mice generated using ESC mEMS681
Resource Files
	Ple167 pEMS1091 Sequence	Jones lab
	Ple167 pEMS1091 MiniPromoter Design	Wasserman lab
	Ple167 pEMS1091 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple167 pEMS1091 Vector Map	Holt lab
	Ple167 pEMS1091 Genotyping Assay	Simpson lab
	Ple167 pEMS1091 ESC description	Simpson lab
	Ple167 pEMS1091 Mouse description	Simpson lab