Ple177-EGFP/cre (Positive)

Ple177-EGFP/cre Description:

The Ple177 promoter construct is derived from the promoter region of the Regulator of G-protein Signaling 16 (RGS16) gene. The Pleiades Promoter Project created a 1302 bp MiniPromoter derived from an upstream candidate regulatory region of the RGS16 gene, along with a subsection of the RGS16 promoter and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1302 (EGFP/cre reporter flanked by minimal frt sites) to make plasmid pEMS1089 with Ple177 driving EGFP/cre. The Ple177-EGFP/cre (pEMS1089) 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 (mEMS761, mEMS762, mEMS763, and mEMS765) were PCR validated. Chimeras were generated using mEMS762.

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

Ple177-EGFP/cre Expression Pattern:

Expression of the Ple177-EGFP/cre results in the rearrangement of the Gt(ROSA)26tm1Sor reporter locus, thus producing strong ßGal activity that is detected widely throughout the brain and eye, with notable expression in the hippocampus, thalamus, forebrain, cortex, brainstem, cerebellum as well as in the retina. In each case, the staining appears mosaic, thus not all cells are ßGal-positive, but a significant subpopulation of cell across many regions are labeled. This mosaicism is emphasized in the labeling of radial columns of cells in the cortex and the retina. A majority of the ßGal-positive cells of the grey matter appear to be neuronal, as they co-label with the neuronal marker NeuN. However, it is apparent that the reporter is not restricted to neurons, but is also detected in cells within the white matter (notably in the corpus callosum, hippocampal fornix and the cerebellar white matter), suggestive of glial labeling as well. EGFP expression was not detected in any region of adult brains, indicating the ßGal expression is an historical marker of expression of the EGFP/cre reporter protein. The broad pattern of reporter expression in the Ple177-EGFP/cre MiniPromoter is very similar, to that seen in the Ple176-EGFP/cre and the Ple178-EGFP/cre strains of mice. Each strain was generated using distinct enhancer elements from the RGS16 gene, but all containing the common basal promoter from this gene as well.

Ple177 MiniPromoter Resources:

Ple177-EGFP/cre
	plasmid pEMS1089 (in backbone pEMS1302)
	ESCs mEMS761, mEMS762, mEMS763, mEMS765
	mice generated using ESC mEMS762
	The mouse strain is available through the Jackson Laboratory: http://jaxmice.jax.org/strain/008877.html
Resource Files
	Ple177 pEMS1089 Sequence	Jones lab
	Ple177 pEMS1089 MiniPromoter Design	Wasserman lab
	Ple177 pEMS1089 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple177 pEMS1089 Vector Map	Holt lab
	Ple177 pEMS1089 Genotyping Assay	Simpson lab

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