The Pleiades Promoter Project

All MiniPromoters

Ple112-EGFP (Positive)

Other constructs for this MiniPromoter: NONE

Other constructs for this Gene: Ple111-EGFP (Positive)


8.4 weeks, N2, Male (entire series)	8.4 weeks, N2, Male (entire series)

Ple112-EGFP Description:

The Ple112 promoter construct is derived from the promoter region of the Hypocretin (HCRT) gene. The Pleiades Promoter Project created a 1295 bp MiniPromoter derived from a subsection of the HCRT promoter and an upstream putative regulatory element. This construct was introduced into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1418 with Ple112 driving EGFP. The Ple112-EGFP (pEMS1418) 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. Four ESC lines (mEMS1722, mEMS1736, mEMS1737, and mEMS1747) were PCR validated. Chimeras were generated using mEMS1736.

Note: a promoterless negative control (pEMS1306) has been generated, as well as a CAG positive controls (pEMS1157, pEMS1277 & pEMS1488).

Ple112 MiniPromoter Resources:

Ple112-EGFP
	plasmid pEMS1418 (in backbone pEMS1306)
	ESCs mEMS1722, mEMS1736, mEMS1737, mEMS1747
	mice generated using ESC mEMS1736
	The mouse strain is available through the Jackson Laboratory: http://jaxmice.jax.org/strain/009594.html
Resource Files
	Ple112 pEMS1418 Sequence	Jones lab
	Ple112 pEMS1418 MiniPromoter Design	Wasserman lab
	Ple112 pEMS1418 Vector NTI File (Requires VectorNTI Sofware)	Holt lab
	Ple112 pEMS1418 Vector Map	Holt lab
	Ple112 pEMS1418 Genotyping Assay	Simpson lab

Publications:

Reference	PubMed ID
Shen HM, Nakamura A, Sugimoto J, Sakumoto N, Oda T, Jinno Y, Okazaki Y. Tissue specificity of methylation and expression of human genes coding for neuropeptides and their receptors, and of a human endogenous retrovirus K family. J Hum Genet. 2006;51(5):440-50. Epub 2006 Mar 17.	16544050
Waleh NS, Apte-Deshpande A, Terao A, Ding J, Kilduff TS. Modulation of the promoter region of prepro-hypocretin by alpha-interferon. Gene. 2001 Jan 10;262(1-2):123-8.	11179675

Sakurai T, Moriguchi T, Furuya K, Kajiwara N, Nakamura T, Yanagisawa M, Goto K. Structure and function of human prepro-orexin gene. J Biol Chem. 1999 Jun 18;274(25):17771-6.	10364220
Moriguchi T, Sakurai T, Takahashi S, Goto K, Yamamoto M. The human prepro-orexin gene regulatory region that activates gene expression in the lateral region and represses it in the medial regions of the hypothalamus. J Biol Chem. 2002 May 10;277(19):16985-92. Epub 2002 Feb 19.	11854267
Zhang S, Lin L, Kaur S, Thankachan S, Blanco-Centurion C, Yanagisawa M, Mignot E, Shiromani PJ. The development of hypocretin (orexin) deficiency in hypocretin/ataxin-3 transgenic rats. Neuroscience. 2007 Aug 10;148(1):34-43. Epub 2007 Jul 6.	17618058

Ple112-EGFP (Positive)

Ple112-EGFP Description:

Ple112-EGFP Expression Pattern:

Ple112 MiniPromoter Resources:

Publications: