Fig. 1
From: rAAV-compatible MiniPromoters for restricted expression in the brain and eye
“Plug and Play” rAAV2 genome plasmid used to clone MiniPromoters (MiniPs) upstream of icre or EmGFP enables high throughput pipeline testing. a Plasmids were generated containing either the icre or the EmGFP reporter. An AsiSI site flanks a removable WPRE. MiniPs were cloned at the MCS using a combination of the four available cut sites. The plasmids are subsequently used to generate single stranded rAAV. b Screening step one, an historical-indirect reporter system. MiniPs drive expression of icre, which in turn recombines the endogenous loxP sites and removes the stop sequence 5ʹ of the lacZ gene, thus driving expression of β-galactosidase from the strong ubiquitous ROSA26 promoter. c Screening step two, a direct reporter system. MiniPs drive direct expression of EmGFP, which can be imaged by epifluorescence or signal amplified using antibodies. bp, base pairs; ITR, inverted terminal repeat; MCS, multiple cloning site; WPRE, woodchuck hepatitis virus post-transcriptional regulatory element