Optogenetics and Transgenic Technology Core

过去的一年是该中心成立核心设施的第一年。我们现在拥有专门从事腺相关病毒（AAV）、慢病毒载体（LV）和犬腺病毒载体（CAV2）的包装、纯化和表征（体外和体内）方法的工作人员。我们经常为NIDA IRP以及其他校内外合作提供载体。这些载体主要用于研究成瘾和神经变性的分子机制。例如，我们已经生产了超过20种不同的载体，用于传递光基因操作神经元回路的视蛋白。我们还开发了几种新的载体来表达突触靶向荧光蛋白、钙敏感荧光蛋白、红外报告蛋白和光敏蛋白，用于光损伤神经元。

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.