Ple103-EGFP/cre (Positive)

Ple103-EGFP/cre Description:

The Ple103 promoter construct is derived from the promoter region of the Huntingtin-Associated Protein 1 (HAP1) gene. The Pleiades Promoter Project created a 877 bp MiniPromoter derived from 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 pEMS1083 with Ple103 driving EGFP/cre. The Ple103-EGFP/cre (pEMS1083) 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 (mEMS675, mEMS676, mEMS689, and mESM1031) were PCR validated. Chimeras were generated using mEMS675.

Note: a promoterless negative control (pEMS1306) has been generated, as well as a CAG positive controls (pEMS1157, pEMS1277 & pEMS1488).

Ple103-EGFP/cre Expression Pattern:

Expression of the Ple103 MiniPromoter driving cre causes rearrangement of the Gt(ROSA)26Sortm1Sor reporter locus, thus producing weak beta-galactosidase activity in a small but consistent population of cells in the brain. These occasional lacZ-positive cells are found throughout the brain; typically these cells are small and found in juxtaposition to blood vessels. Rare large lacZ-positive cells are detected in neuropil of the forebrain and midbrain. 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.

Ple103 MiniPromoter Resources:

Ple103-EGFP/cre
	plasmid pEMS1083 (in backbone pEMS1302)
	ESCs mEMS675, mEMS676, mEMS689, mEMS1031
	mice generated using ESC mEMS675
	The mouse strain is available through the Jackson Laboratory: http://jaxmice.jax.org/strain/009114.html
Resource Files
	Ple103 pEMS1083 Sequence	Jones lab
	Ple103 pEMS1083 MiniPromoter Design	Wasserman lab
	Ple103 pEMS1083 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	Ple103 pEMS1083 Vector Map	Holt lab
	Ple103 pEMS1083 Genotyping Assay	Simpson lab
	Ple103 pEMS1083 ESC description	Simpson lab
	Ple103 pEMS1083 Mouse description	Simpson lab

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