For cell type-specific targeting and to capitalize on the large number of Cre driver lines with the flexible virus injection/fiberoptic approach, we have developed a tool we call DIO-AAV (doublefloxed inverse orf) AAV.
| pAAV-Ef1a-DIO-hChR2(H134R)-EYFP-WPRE-pA | [ Vector Map ] |
| pAAV-Ef1a-DIO-hChR2(H134R)-mCherry-WPRE-pA | [ Vector Map ] |
| pAAV-Ef1a-DIO-eNpHR-EYFP-WPRE-pA | [ Vector Map ] |
Optical Excitation:
Channelrhodopsin-2 (ChR2)
There are two versions of the ChR2 sequence, one containing the wildtype sequence and another containing condons optimized for mammalian expression (hChR2). In-frame fusions to mCherry or EYFP are available to make visualization of ChR2-expressing cells easier. ChR2 and XFP are fused via a NotI site. The linker is GCGGCCGCC.
ChR2-XFPs are available either in a standard mammalian expression vector containing the CMV promoter or in lentiviral expression vectors under the control of the ubiquitous EF-1a or the neuron-specific CaMKIIa or human Synapsin I promoters. The structure of the lentivirus is shown below.
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| pcDNA3.1/hChR2-mCherry | [ Vector Map ] |
| pcDNA3.1/hChR2-EYFP | [ Vector Map ] |
| pcDNA3.1/hChR2(H134R)-EYFP | [ Vector Map ] |
| pAAV-CaMKIIa-hChR2(H134R)-EYFP | [ Vector Map ] |
| pAAV-CaMKIIa-hChR2(H134R)-mCherry | [ Vector Map ] |
| pAAV-hSyn-hChR2(H134R)-EYFP | [ Vector Map ] |
| pAAV-hSyn-hChR2(H134R)-mCherry | [ Vector Map ] |
| pAAV-hSyn-hChR2(H134R)-WPRE | [ Vector Map ] |
| pAAV-hSyn-hChR2(H134R)-p2A-WPRE | [ Vector Map ] |
| pAAV-mThy1-hChr2(H134R)-eYFP-WPRE | [ Vector Map ] |
| pAAV-hThy1-hChR2(H134R)-eYFP-WPRE | [ Vector Map ] |
| pAAV-GFAP-hChR2(H134R)-EYFP | [ Vector Map ] |
| pAAV-GFAP-hChR2(H134R)-mCherry | [ Vector Map ] |
| pAAV-EF1a-hChR2(H134R)-EYFP | [ Vector Map ] |
| pLenti-EF1a-hChR2-EYFP-WPRE (a.k.a. pLECYT) | [ Vector Map ] |
| pLenti-EF1a-hChR2(H134R)-EYFP-WPRE (a.k.a. pLECYT) | [ Vector Map ] |
| pLenti-CaMKIIa-hChR2-mCherry-WPRE | [ Vector Map ] |
| pLenti-CaMKIIa-hChR2-EYFP-WPRE | [ Vector Map ] |
| pLenti-Hcrt-hChR2(H134R)-EYFP | [ Vector Map ] |
| pLenti-Synapsin-hChR2(H134R)-EYFP-WPRE | [ Vector Map ] |
| pLenti-mThy1-hChR2(H134R)-eYFP | [ Vector Map ] |
| pLenti-hThy1-hChR2(H134R)-eYFP | [ Vector Map ] |