This past year marks the first year as an established core facility. We now have staff who are specilized in the methods to package, purify and characterize (in vitro and in vivo) both adeno-associated viral (AAV), lentiviral vectors (LV)and Canine adenoviral vectors (CAV2). 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, calcium-sensitive fluorescent proteins, infrared reporter protein and photosensitizing proteins for lesioning neurons with light. We have successfully created but not yet published several transgenic rat lines including a conditional-mCherry fluorescent reporter rat that will only express the mCherry fluorescent protein where Cre recombinase is present. This rat has been crossed with our DAT-Cre rat to verify expression of Cre in dopaminergic neurons. We also have a transgenic rat line expressing 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. We have additional Cre-driver rats in early stages of characterization as well as other reporter rats. All of our characterized and published vectors are available for intramural and extramural collaborators.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Scientific Cores Intramural Research (ZIC)
Project #
1ZICDA000555-05
Application #
8736963
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2013
Total Cost
$460,579
Indirect Cost
Name
National Institute on Drug Abuse
Department
Type
DUNS #
City
State
Country
Zip Code
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
Lee, Chun-Ting; Chen, Jia; Kindberg, Abigail A et al. (2016) CYP3A5 Mediates Effects of Cocaine on Human Neocorticogenesis: Studies using an In Vitro 3D Self-Organized hPSC Model with a Single Cortex-Like Unit. Neuropsychopharmacology :
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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