The Pleiades Promoter Project

All MiniPromoters

Ple80-Cre/EGFP (Negative)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple77-Cre/EGFP (Negative), Ple78-EGFP (Negative), Ple79-Cre/EGFP (Negative), Ple79-EGFP (Negative)

8.9 weeks, N2, Male
(entire series)

Ple80-EGFP/cre Description:

This Ple80 promoter construct is derived from the promoter region of the gamma-aminobutyric acid receptor, subunit alpha 6 (GABRA6) gene. The Pleiades Promoter Project created a 2184 bp MiniPromoter derived from two upstream putative regulatory regions along with a section of the promoter and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1040 (cre/EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1060 with Ple80 driving cre/EGFP. The Ple80-cre/EGFP (pEMS1060) 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) mEMS21. Four ESC lines (mEMS503, mEMS505, mEMS510, and mEMS512) were PCR validated. Chimeras were generated using mEMS505.

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

Ple80-EGFP/cre Expression Pattern:

MiniPromoter Ple80 was assayed for LacZ and EGFP from pEMS1040 backbone. No xGal reaction was detected in numerous brain regions and retina. 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 and retina (brightfield).

Ple80 MiniPromoter Resources:

Ple80-cre/EGFP
	plasmid pEMS1060 (in backbone pEMS1040)
	ESCs mEMS503, mEMS505, mEMS510, and mEMS512
	mice generated using ESC mEMS505
Resource Files
	Ple80 pEMS1060 Sequence	Jones lab
	Ple80 pEMS1060 MiniPromoter Design	Wasserman lab
	Ple80 pEMS1060 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple80 pEMS1060 Vector Map	Holt lab
	Ple80 pEMS1060 Vector Map (Requires VectorNTI Sofware)	Holt lab
	Ple80 pEMS1060 Genotyping Assay (Requires VectorNTI Sofware)	Holt lab
	Ple80 pEMS1060 Genotyping Assay	Simpson lab
	Ple80 pEMS1060 ESC description	Simpson lab
	Ple80 pEMS1060 Mouse description	Simpson lab

Publications:

Reference	PubMed ID
Jones A, Korpi ER, McKernan RM, Pelz R, Nusser Z, Makela R, Mellor JR, Pollard S, Bahn S, Stephenson FA, Randall AD, Sieghart W, Somogyi P, Smith AJ, Wisden W. Ligand-gated ion channel subunit partnerships: GABAA receptor alpha6 subunit gene inactivation inhibits delta subunit expression. J Neurosci. 1997 Feb 15;17(4):1350-62.	9006978
Bahn S, Jones A, Wisden W. Directing gene expression to cerebellar granule cells using gamma-aminobutyric acid type A receptor alpha6 subunit transgenes. Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9417-21.	9256497
Wang W, Stock RE, Gronostajski RM, Wong YW, Schachner M, Kilpatrick DL. A role for nuclear factor I in the intrinsic control of cerebellar granule neuron gene expression. J Biol Chem. 2004 Dec 17;279(51):53491-7. Epub 2004 Oct 1.	15466411
McLean PJ, Shpektor D, Bandyopadhyay S, Russek SJ, Farb DH. A minimal promoter for the GABA(A) receptor alpha6-subunit gene controls tissue specificity. J Neurochem. 2000 May;74(5):1858-69.	10800928
Jones A, Bahn S, Grant AL, Kohler M, Wisden W. Characterization of a cerebellar granule cell-specific gene encoding the gamma-aminobutyric acid type A receptor alpha 6 subunit. J Neurochem. 1996 Sep;67(3):907-16.	8752095
Zheng T, Santi MR, Bovolin P, Marlier LN, Grayson DR. Developmental expression of the alpha 6 GABAA receptor subunit mRNA occurs only after cerebellar granule cell migration. Brain Res Dev Brain Res. 1993 Sep 17;75(1):91-103.	8222213
Wang W, Shin Y, Shi M, Kilpatrick DL. Temporal Control of a Dendritogenesis-Linked Gene via Rest-Dependent Regulation of Nuclear Factor I Occupancy. Mol Biol Cell. 2011 Jan 26; [Epub ahead of print]	21270437