The Pleiades Promoter Project

All MiniPromoters

Ple77-Cre/EGFP (Negative)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple78-EGFP (Negative), Ple79-Cre/EGFP (Negative), Ple79-EGFP (Negative), Ple80-Cre/EGFP (Negative)

9 weeks, N2, Male
(entire series)

Ple77-EGFP/cre Description:

This Ple77 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 668 bp MiniPromoter derived from a portion of the promoter of the GABRA6 gene introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1040 (Cre/EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1057 with Ple77 driving cre/EGFP. The Ple77-cre/EGFP (pEMS1057) 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 (mEMS554, mEMS555, mEMS563, and mEMS567) were PCR validated. Chimeras were generated using mEMS554.

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

Ple77-EGFP/cre Expression Pattern:

MiniPromoter Ple77 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).

Ple77 MiniPromoter Resources:

Ple77-cre/EGFP
	plasmid pEMS1057 (in backbone pEMS1040)
	ESCs mEMS554, mEMS555, mEMS563, and mEMS567
	mice were generated using ESC mEMS554
Resource Files
	Ple77 pEMS1057 Sequence	Jones lab
	Ple77 pEMS1057 MiniPromoter Design	Wasserman lab
	Ple77 pEMS1057 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple77 pEMS1057 Vector Map	Holt lab
	Ple77 pEMS1057 Genotyping Assay	Simpson lab
	Ple77 pEMS1057 ESC description	Simpson lab
	Ple77 pEMS1057 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