The Pleiades Promoter Project

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All MiniPromoters

Ple66-EGFP (Negative)

Other constructs for this MiniPromoter: Ple66-lacZ (Negative)

Other constructs for this Gene: Ple67-EGFP (Positive), Ple67-lacZ (Positive)

7.4 weeks, N2, Male
(entire series)

Ple66-EGFP Description:

The Ple66 promoter construct is derived from the promoter region of the FEV (ETS oncogene family) (FEV) gene. The Pleiades Promoter Project created a 1607 bp MiniPromoter derived from a subsection of the FEV promoter and a downstream putative regulatory element and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1400 with Ple66 driving EGFP. The Ple66-EGFP (pEMS1400) 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. Three ESC lines (mEMS2792, mEMS2793, and mEMS2794) were PCR validated. Chimeras were generated using mEMS2792.

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

Ple66-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).

Ple66 MiniPromoter Resources:

Ple66-EGFP
	plasmid pEMS1400 (in backbone pEMS1306)
	ESCs mEMS2792, mEMS2793, mEMS2794
	mice generated using ESC mEMS2792
Resource Files
	Ple66 pEMS1400 Sequence	Jones lab
	Ple66 pEMS1400 MiniPromoter Design	Wasserman lab
	Ple66 pEMS1400 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple66 pEMS1400 Vector Map	Holt lab
	Ple66 pEMS1400 Genotyping Assay (Requires VectorNTI Sofware)	Holt lab

Publications:

Reference	PubMed ID
Scott MM, Krueger KC, Deneris ES. A differentially autoregulated Pet-1 enhancer region is a critical target of the transcriptional cascade that governs serotonin neuron development. J Neurosci. 2005 Mar 9;25(10):2628-36.	15758173
Scott MM, Wylie CJ, Lerch JK, Murphy R, Lobur K, Herlitze S, Jiang W, Conlon RA, Strowbridge BW, Deneris ES. A genetic approach to access serotonin neurons for in vivo and in vitro studies. Proc Natl Acad Sci U S A. 2005 Nov 8;102(45):16472-7. Epub 2005 Oct 26.	16251278
Pfaar H, von Holst A, Vogt Weisenhorn DM, Brodski C, Guimera J, Wurst W. mPet-1, a mouse ETS-domain transcription factor, is expressed in central serotonergic neurons. Dev Genes Evol. 2002 Feb;212(1):43-6. Epub 2002 Jan 31.	11875656
Maurer P, Rorive S, de Kerchove d'Exaerde A, Schiffmann SN, Salmon I, de Launoit Y. The Ets transcription factor Fev is specifically expressed in the human central serotonergic neurons. Neurosci Lett. 2004 Mar 11;357(3):215-8.	15003288
Iyo AH, Porter B, Deneris ES, Austin MC. Regional distribution and cellular localization of the ETS-domain transcription factor, FEV, mRNA in the human postmortem brain. Synapse. 2005 Sep 15;57(4):223-8.	15986391
Krueger KC, Deneris ES. Serotonergic Transcription of Human FEV Reveals Direct GATA Factor Interactions and Fate of Pet-1-Deficient Serotonin Neuron Precursors. J Neurosci. 2008 Nov 26;28(48):12748-58.	19036967