The Pleiades Promoter Project

All MiniPromoters

Ple150-EGFP (Negative)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple148-EGFP (Negative), Ple151-EGFP (Positive)

Ple150-EGFP Description:

The Ple150 promoter construct is derived from the promoter region of the oligodendrocyte transcription factor 1 (OLIG1) gene. The Pleiades Promoter Project designed a 3194 bp MiniPromoter derived from three upstream putative regulatory elements, along with a subsection of the OLIG1 promoter, and introduced them into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1171 with Ple150 driving EGFP. The Ple150-EGFP (pEMS1171) 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 (mEMS1605, mEMS1607, mEMS1616, and mEMS1617) were PCR validated. Chimeras were generated using mEMS1607.

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

Ple150-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).

Ple150 MiniPromoter Resources:

Ple150-EGFP
	plasmid pEMS1171 (in backbone pEMS1306)
	ESCs mEMS1605, mEMS1607, mEMS1616, mEMS1617
	mice generated using ESC mEMS1607
Resource Files
	Ple150 pEMS1171 MiniPromoter Design	Wasserman lab
	Ple150 pEMS1171 Genotyping Assay	Simpson lab
	Ple151 pEMS1171 Mouse description	Simpson lab

Publications:

Reference	PubMed ID
Lu QR, Yuk D, Alberta JA, Zhu Z, Pawlitzky I, Chan J, McMahon AP, Stiles CD, Rowitch DH. Sonic hedgehog--regulated oligodendrocyte lineage genes encoding bHLH proteins in the mammalian central nervous system. Neuron. 2000 Feb;25(2):317-29.	10719888
Qi Y, Cai J, Wu Y, Wu R, Lee J, Fu H, Rao M, Sussel L, Rubenstein J, Qiu M. Control of oligodendrocyte differentiation by the Nkx2.2 homeodomain transcription factor. Development. 2001 Jul;128(14):2723-33.	11526078
Gong X, Lin T, Sun Z, Fu M, Zuo H, Xie Z. Olig1 is downregulated in oligodendrocyte progenitor cell differentiation. Neuroreport. 2008 Aug 6;19(12):1203-1207.	18628665
Ikushima H, Komuro A, Isogaya K, Shinozaki M, Hellman U, Miyazawa K, Miyazono K. An Id-like molecule, HHM, is a synexpression group-restricted regulator of TGF-beta signalling. EMBO J. 2008 Oct 16; [Epub ahead of print]	18923419