This past year marks the initial establishment of the Optogenetics and Transgenic Technology Core. We have hired a group of people who are now trained in the methods to package, purify and characterize (in vitro and in vivo) both adeno-associated viral (AAV) vectors and lentiviral vectors (LV). We routinely generate vectors both for the NIDA IRP as well as other intramural and extramural collaborations. The vectors are designed primarily for studies focused on molecular mechanisms of addiction and neurodegeneration. For example, we have produced over 20 different vectors for delivery of the opsins for optogenetic manipulation of neuronal circuits. We have also been developing several novel vectors to express synaptically targeted fluorescent proteins, voltage sensitive fluorescent proteins, calcium-sensitive fluorescent proteins, infrared reporter protein and photosensitizing proteins for lesioning neurons with light. We have our first generation of transgenic rats to be characterized. We have a conditional-mCherry reporter rat that will only express the mCherry fluorescent protein where Cre recombinase is present. This rat can be used to test Cre expressing viral vectors or crossed with our other cell specific, Cre expressing rats to examine the specificity of Cre expression. We also have pups for our animals expressing the Tet repressor protein. The Tet repressor protein can bind to specific DNA elements (Tet operator) to prevent gene expression. By giving animals tetracycline or an analog we can cause the repressor to come off the DNA and allow gene expression. This will be used in combination with viruses or other rats that containing the Tet operator to allow us to activate gene expression. Constructs have also been injected into fertilized eggs for a microglia specific expression of green fluorescent protein, the dopamine transporter promoter expressing Cre recombinase and the Fos promoter, a marker of neuronal activity, expressing Cre recombinase. These animals will be characterized over the next several months. All of our characterized and published vectors are available for intramural and extramural collaborators.

National Institute of Health (NIH)
National Institute on Drug Abuse (NIDA)
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Hamra, F Kent; Richie, Christopher T; Harvey, Brandon K (2017) Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production. Transgenic Res 26:477-489
Wallace, Veronica J; Cimbro, Raffaello; Rubio, F Javier et al. (2017) Neurons Internalize Functionalized Micron-Sized Silicon Dioxide Microspheres. Cell Mol Neurobiol 37:1487-1499
Sharpe, Melissa J; Marchant, Nathan J; Whitaker, Leslie R et al. (2017) Lateral Hypothalamic GABAergic Neurons Encode Reward Predictions that Are Relayed to the Ventral Tegmental Area to Regulate Learning. Curr Biol 27:2089-2100.e5
Howard, Douglas B; Harvey, Brandon K (2017) Assaying the Stability and Inactivation of AAV Serotype 1 Vectors. Hum Gene Ther Methods 28:39-48
Wires, Emily S; Henderson, Mark J; Yan, Xiaokang et al. (2017) Longitudinal monitoring of Gaussia and Nano luciferase activities to concurrently assess ER calcium homeostasis and ER stress in vivo. PLoS One 12:e0175481
Richie, Christopher T; Whitaker, Leslie R; Whitaker, Keith W et al. (2017) Near-infrared fluorescent protein iRFP713 as a reporter protein for optogenetic vectors, a transgenic Cre-reporter rat, and other neuronal studies. J Neurosci Methods 284:1-14
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Kuo, Chi-Chung; Shen, Hui; Harvey, Brandon K et al. (2016) Differential modulation of methamphetamine-mediated behavioral sensitization by overexpression of Mu opioid receptors in nucleus accumbens and ventral tegmental area. Psychopharmacology (Berl) 233:661-72
Marchant, Nathan J; Whitaker, Leslie R; Bossert, Jennifer M et al. (2016) Behavioral and Physiological Effects of a Novel Kappa-Opioid Receptor-Based DREADD in Rats. Neuropsychopharmacology 41:402-9
Marchant, Nathan J; Campbell, Erin J; Whitaker, Leslie R et al. (2016) Role of Ventral Subiculum in Context-Induced Relapse to Alcohol Seeking after Punishment-Imposed Abstinence. J Neurosci 36:3281-94

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