The Pleiades Promoter Project

All MiniPromoters

Ple175-EGFP (Negative)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple176-EGFP/cre (Positive), Ple177-EGFP/cre (Positive), Ple178-EGFP/cre (Positive), Ple176-EGFP (Negative), Ple177-EGFP (Negative), Ple178-EGFP (Negative), Ple178-lacZ (Negative)

Ple175-EGFP Description:

The Ple175 promoter construct is derived from the promoter region of the Regulator of G-protein Signaling 16 (RGS16) gene. The Pleiades Promoter Project created a 581 bp MiniPromoter derived from a subsection of the RGS16 promoter and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1363 with Ple175 driving EGFP. The Ple175-EGFP (pEMS1363) 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) mEMS1204.03A. Four ESC lines (mEMS1654, mEMS1657, mEMS1658, and mEMS1659) were PCR validated. Chimeras were generated using mEMS1654.

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

Ple175-EGFP Expression Pattern:

No EGFP expression was detected above background in native fluorescence under epi or confocal microscopy. Anti GFP immunostaining (rabbit polyclonal, Invitrogen) did not detect specific GFP expression above background across numerous brain regions (brightfield).

Ple175 MiniPromoter Resources:

Ple175-EGFP
	plasmid pEMS1363 (in backbone pEMS1306)
	ESCs mEMS1654, mEMS1657, mEMS1658, mEMS1659
	mice generated using ESC mEMS1654
Resource Files
	Ple175 pEMS1363 Sequence	Jones lab
	Ple175 pEMS1363 MiniPromoter Design	Wasserman lab
	Ple175 pEMS1363 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple175 pEMS1363 Vector Map	Holt lab
	Ple175 pEMS1363 Genotyping Assay	Simpson lab

Publications:

Reference	PubMed ID
Chen C, Zheng B, Han J, Lin SC. Characterization of a novel mammalian RGS protein that binds to Galpha proteins and inhibits pheromone signaling in yeast. J Biol Chem. 1997 Mar 28;272(13):8679-85.	9079700
Panetta R, Guo Y, Magder S, Greenwood MT. Regulators of G-protein signaling (RGS) 1 and 16 are induced in response to bacterial lipopolysaccharide and stimulate c-fos promoter expression. Biochem Biophys Res Commun. 1999 Jun 16;259(3):550-6.	10364456
Fong CW, Zhang Y, Neo SY, Lin SC. Specific induction of RGS16 (regulator of G-protein signalling 16) mRNA by protein kinase C in CEM leukaemia cells is mediated via tumour necrosis factor alpha in a calcium-sensitive manner. Biochem J. 2000 Dec 15;352 Pt 3:747-53.	11104682
Grafstein-Dunn E, Young KH, Cockett MI, Khawaja XZ. Regional distribution of regulators of G-protein signaling (RGS) 1, 2, 13, 14, 16, and GAIP messenger ribonucleic acids by in situ hybridization in rat brain. Brain Res Mol Brain Res. 2001 Mar 31;88(1-2):113-23.	11295237
Berthebaud M, Riviere C, Jarrier P, Foudi A, Zhang Y, Compagno D, Galy A, Vainchenker W, Louache F. RGS16 is a negative regulator of SDF-1-CXCR4 signaling in megakaryocytes. Blood. 2005 Nov 1;106(9):2962-8. Epub 2005 Jul 5.	15998835
Stuebe S, Wieland T, Kraemer E, Stritzky A, Schroeder D, Seekamp S, Vogt A, Chen CK, Patten M. Sphingosine-1-phosphate and endothelin-1 induce the expression of rgs16 protein in cardiac myocytes by transcriptional activation of the rgs16 gene. Naunyn Schmiedebergs Arch Pharmacol. 2008 Jan;376(5):363-73. Epub 2007 Nov 29.	18046543
Wiechec E, Overgaard J, Hansen LL. A fragile site within the HPC1 region at 1q25.3 affecting RGS16, RGSL1, and RGSL2 in human breast carcinomas. Genes Chromosomes Cancer. 2008 Jun 2; [Epub ahead of print]	18521847