DAT-eArchT3.0-EGFP BAC transgenic mice have the mouse dopamine active transporter (Slc6a3) promoter/enhancer regions directing expression of the optimized archeorhodopsin eArchT3.0 and EGFP to dopaminergic neuronal populations (see below for detailed description of eArchT3.0). The donating investigator reports that prior to illumination with yellow-green light, DAT-eArchT3.0-EGFP mice exhibit robust and bright EGFP fluorescence and there is no hyperpolarization (although eArchT3.0 expression is observable by immunochemical detection via the P2A fragment at the eArchT3.0 C-terminus). Illumination of eArchT3.0-expressing neurons with yellow-green light leads to reversible photoinhibition of action potential firing/neural activity in these cells, while EGFP fluorescence remains robust and bright. eArchT3.0 or EGFP expression in tissues other than brain has not yet been characterized (June 2018). Mice hemizygous for the DAT-eArchT3.0-EGFP BAC transgene are viable and fertile with no reported gross physical or behavioral abnormalities. It has not been attempted to make this strain homozygous to date (June 2018). The bacterial opsins are retinal-binding proteins that combine a light-sensitive domain with an ion channel or pump; providing light-dependent ion transport, membrane potential alteration and sensory functions to bacteria. eArchT3.0 is a mammalian codon-optimized, yellow-green light-driven (~520-560 nm), outward proton pump that causes hyperpolarization and prevents action potentials [<a href="https://www.ncbi.nlm.nih.gov/pubmed/22179551">Mattis et al. 2012 Nat Methods 9:159</a>]. It is a variant of the Halorubrum strain TP009-derived Archaerhodopsin-3 (ArchT) that was further modified to have a neurite/membrane-trafficking signal (TS) and endoplasmic reticulum (ER) exporting sequence. These improvements result in enhanced membrane localization and reduced intracellular accumulation - and subsequently much larger photocurrents (two-fold increase) compared to ArchT or Arch.

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DAT-eArchT3.0-EGFP BAC transgenic mice allow optogenetic silencing of dopaminergic neurons using eArchT3.0 - a mammalian codon-optimized, outward proton pump that causes hyperpolarization and prevents action potentials in response to yellow-green light (~520-560 nm), with enhanced membrane localization and reduced intracellular accumulation, and subsequently much larger photocurrents (two-fold increase) compared to ArchT or Arch.

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