CAG-EGFP (pEMS1157)

CAG-EGFP Description:

The CAG promoter is generally reported to have a wide expression throughout cell types and development (resources). To act as a positive control and to generate new ubiquitously expressing resources, the Pleiades Promoter Project isolated the CAG promoter from the InvivoGen pDRIVE-CAG vector and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1306 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1157 with CAG driving EGFP. The CAG-EGFP (pEMS1157) was introduced by homologous recombination as single copy insertion into the Hprt1b-m3 locus in the X Chromosome of mouse embryonic stem cells (ESCs) mEMS1204. The resulting PCR validated four ESC lines were mEMS1561, mEMS1589, mEMS1590, and mEMS1600. Chimeras were generated using mEMS1589, mEMS1590, and mEMS1600 and the former two cell lines were used to generate the images below showing ubiquitous expression as expected of this positive control.

In a parallel experiment, also to generate a positive control, the CAG promoter was introduced into the MCS of Pleiades expression vector pEMS1277 (EGFP reporter flanked by full frt sites) to make plasmid pEMS1277 with CAG driving EGFP.

Note: Promoterless negative controls (pEMS1302, pEMS1306, pEMS1313) have also been generated.

CAG MiniPromoter Resources:

CAG-EGFP
	plasmid pEMS1157 (in backbone pEMS1306)
	ESCs mEMS1561, mEMS1589, mEMS1590, and mEMS1600
	chimeras generated using ESCs mEMS1589, mEMS1590, and mEMS1600
Resource Files
	CAG Sequence	Jones lab
	CAG MiniPromoter Design	Wasserman lab
	CAG pEMS1157 Vector NTI File  (Requires VectorNTI Sofware)	Holt lab
	CAG pEMS1157 Vector Map	Holt lab
	CAG pEMS1157 Genotyping Assay	Simpson lab
	CAG ESC description	Simpson lab
	CAG Mouse description	Simpson lab

Publications: