In this webinar, CLOVER Director Tim Miles introduces viewers to established and emerging engineered systemic AAVs from the Gradinaru lab and others, highlighting each capsid's comparative strengths and key variables for successful implementation.
The question and answer session is hosted by Jason Nasse, Senior Scientist-Viral Communications at Addgene, and includes Gradinaru lab graduate students David Goertson, Xinhong Chen, and Miggy Chuapoco.
Answers to additional questions we didn't get to in the live event are available here.
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0:00 - Intro
2:10 - Main Presentation
20:18 - Q&A
- 20:57 - What advice would you give to someone just starting to use systemic AAVs in their experiments?
- 23:20 - Did Adenoviruses and AAVs co-evolve? Is directed evolution possible for Adenoviral vectors?
- 24:18 - Is retro-orbital or tail vein more reproducible in your hands?
- 25:28 - Do you have any plans to optimize capsids to target microglia?
- 26:29 - If AAV.CAP-B10 is better for Glutamatergic neurons, what about GABAergic neurons?
- 27:54 - Which serotype of AAV are recommended for neuro-muscular junction?
- 28:59 - Your talk focused on mice, but for those of us that use rats can you provide any advice? If it works in mice, can we assume they will work in Sprague rats or Long Evans rats?
- 31:40 - Can I inject a PHP.eB AAV and another PHP (e.g., PHP.S) a few weeks later to the same mouse?
- 33:20 - I wonder what's the best route for administering AAVs to target neonatal and adult marmoset brain? Is there a plan to optimize based on existing tools?
- 34:27 - Have you ever noticed any baseline effects on mouse behavior with PHP.eB AAVs with empty vectors?
- 36:18 - Given that genetic variability affects expression in marmosets, which are much more variable than mice, is expression of cargo very variable for different animals? Or is it consistent?
- 37:45 - Are there systemic AAVs suitable for ventricular injection that you would recommend (for mice)?
- 38:53 - What constrains the amount of AAV that can be injected?
- 41:48 - Has your group or others looked at the role of post-translational modifications of capsids for viral tropism?
- 42:49 - Are there brain region differences in the tropism of CAP-B10 AAVs?
- 43:55 - How can we control MOI in vivo? Did you ever check how many viruses are infecting one cell?
- 45:56 - Is it possible to target sensory vs. motor peripheral neurons?
- 46:44 - How do you get rid of empty the capsids in your preps?
- 48:41 - Does transducing astrocytes activate them?
- 50:16 - Does the lack of BBB or leaky BBB influence the expression of PHP.eB?
- 51:44 - Is it possible to conduct a workshop in the future to get people hands experience on learning how to make the virus all the way to injection in mice/rats and check expression?
53:11 - Outro