The Pleiades Promoter Project

All MiniPromoters

Ple54-EGFP (Positive)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple53-EGFP (Positive), Ple53-lacZ (Positive), Ple55-EGFP (Positive), Ple55-lacZ (Positive)


12 weeks, N2, Male (entire series)	(entire series)

Ple54-EGFP Description:

The Ple54 promoter construct is derived from the promoter region of the Doublecortin (DCX) gene. The Pleiades Promoter Project created a 3687 bp MiniPromoter derived from a subsection of the DCX promoter and two intronic candidate regulatory elements. This construct was introduced into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1200 with Ple54 driving EGFP. The Ple54-EGFP (pEMS1200) 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. Four ESC lines (mEMS1927, mEMS1928, mEMS1930, and mEMS1937) were PCR validated. Chimeras were generated using mEMS1927.

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

Ple54-EGFP Expression Pattern:

In Ple54-EGFP MiniPromoter mice detection of EGFP is strongest in soma of cells in the neurogenic regions of the adult brain: the subgranular zone of the dentate gyrus as well as the forebrain subventricular zone (rostral migratory stream and olfactory bulb). Additionally, EGFP-positive cell bodies are detected in the neuropil of the cortex, hippocampus and brainstem. Double label immunocytochemistry shows that the EGFP-positive cells of the forebrain subventricular zone and the rostral migratory stream co-express Doublecortin (the product of the DCX gene from which the Ple54-EGFP MiniPromoter was derived).

Ple54 MiniPromoter Resources:

Ple54-EGFP
	plasmid pEMS1200 (in backbone pEMS1306)
	ESCs mEMS1927, mEMS1928, mEMS1930, mEMS1937
	mice generated using ESC mEMS1927
Resource Files
	Ple54 pEMS1200 Sequence	Jones lab
	Ple54 pEMS1200 MiniPromoter Design	Wasserman lab
	Ple54 pEMS1200 Vector NTI File	Holt lab
	Ple54 pEMS1200 Vector Map	Holt lab
	Ple54 pEMS1200 Genotyping Assay	Simpson lab

Publications:

Reference	PubMed ID
Karl C, Couillard-Despres S, Prang P, Munding M, Kilb W, Brigadski T, Plotz S, Mages W, Luhmann H, Winkler J, Bogdahn U, Aigner L. Neuronal precursor-specific activity of a human doublecortin regulatory sequence. J Neurochem. 2005 Jan;92(2):264-82.	15663475
Couillard-Despres S, Winner B, Karl C, Lindemann G, Schmid P, Aigner R, Laemke J, Bogdahn U, Winkler J, Bischofberger J, Aigner L. Targeted transgene expression in neuronal precursors: watching young neurons in the old brain. Eur J Neurosci. 2006 Sep;24(6):1535-45.	17004917
Wang X, Mao X, Xie L, Greenberg DA, Jin K. Involvement of Notch1 signaling in neurogenesis in the subventricular zone of normal and ischemic rat brain in vivo. J Cereb Blood Flow Metab. 2009 Jun 17; [Epub ahead of print]	19536070
Piens M, Muller M, Bodson M, Baudouin G, Plumier JC. A short upstream promoter region mediates transcriptional regulation of the mouse doublecortin gene in differentiating neurons. BMC Neurosci. 2010 May 28;11(1):64 [Epub ahead of print]	20509865