P02 Platform for recombinant adeno-associated- , lenti-, and modified rabies virus-based technologies and advanced light microscopy
This project will serve as a platform to generate and purify recombinant mainly adeno-associated viral particles that will be used in multiple projects for in vivo applications. In recent years, viral vectors have emerged as a general approach for in vivo expression because of their versatility, efficiency, and relative ease of use. During the last funding period P02 has successfully purified over 294 different custom viruses and provided them to multiple groups within the SFB (Fuhrmann, Remy, Beck, Mody, Blaess, Ewell, Bradke, Dietrich, Henneberger, Schwarz, Schoch, van Loo/Becker). These included viral expression vectors that were purchased or obtained from other labs by the individual investigators. However, the majority of the viral vectors were cloned by the viral vector platform (around 104). Therefore, the repertoire of the platform now includes viral vectors with multiple different promoters (for example CMV, CAG, CBA, EF1a, synapsin, CamKII, GFAP, Cav3.2, arc), various reporters (fluorescent proteins ranging from blue to infrared), sensors (Ca2+-, voltage-sensors), light-activated channels and markers (for example for pre- and postsynaptic). P02 will continue to provide the individual subprojects with the required purified rAAV- and lenti-viral particles, and generate custom viral vectors as needed. In addition, P02 will provide virus-based fluorescent labeling of functional neuronal circuits (i.e. with recombinant rabies viruses) and subsequent analysis with light microscopic techniques (3D confocal microscopy, light sheet fluorescent microscopy (LSFM), expansion light sheet fluorescent microscopy (ELSFM)).