The Pleiades Promoter Project

All MiniPromoters

Ple53-EGFP (Positive)

Other constructs for this MiniPromoter: Ple53-lacZ (Positive)

Other constructs for this Gene: Ple54-EGFP (Positive), Ple55-EGFP (Positive), Ple55-lacZ (Positive)


7.9 weeks, N2, Male (entire series)	7.9 weeks, N2, Male (entire series)

Ple53-EGFP Description:

The Ple53 promoter construct is derived from the promoter region of the Doublecortin (DCX) gene. The Pleiades Promoter Project created a 3520 bp MiniPromoter derived from a subsection of the DCX promoter. 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 pEMS1892 with Ple53 driving EGFP. The Ple53-EGFP (pEMS1199) 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 (mEMS1886, mEMS1892, mEMS1894, and mEMS1913) were PCR validated. Chimeras were generated using mEMS1892.

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

Ple53-EGFP Expression Pattern:

EGFP-immunoreactivity is detected most strongly in the neurogenic cells of the rostral migratory stream (RMS) similar to the RMS expression seen with the Ple54-EGFP MiniPromoter. In contrast to the expression pattern of Ple54-EGFP, expression from the Ple53-EGFP MiniPromoter is weaker but more specific; being not found in cells of the cortex, hippocampus, or brainstem.

Ple53 MiniPromoter Resources:

Ple53-EGFP
	plasmid pEMS1199 (in backbone pEMS1306)
	ESCs mEMS1886, mEM1892, mEMS1894, mEMS1913
	mice generated using ESC mEMS1892
Resource Files
	Ple53 pEMS1199 Sequence	Jones lab
	Ple53 pEMS1199 MiniPromoter Design	Wasserman lab
	Ple53 pEMS1199 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple53 pEMS1199 Vector Map	Holt lab
	Ple53 pEMS1199 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