Navigating an Expanding Landscape for Engineered Systemic AAVs - with Dr. Tim Miles (Webinar)
In this webinar, Dr. Tim Miles, director of the CLOVER Center at Caltech, will introduce 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.
As director of the CLOVER Center at Caltech, Dr. Tim Miles works to further develop engineered AAV technologies and facilitate the broad adoption of novel AAV tools. This has included collaborating with Prof. Viviana Gradinaru to help develop the Multiplexed-CREATE methodology for AAV capsid engineering and providing collaborative advice, training, and AAV reagents to scores of academic labs each year.
Additional Q&A Questions/Answers on the Addgene Blog: https://blog.addgene.org/aav-qa-with-...
https://www.clover.caltech.edu
https://datahub.addgene.org/
Featuring:
Dr. Tim Miles, Director of the CLOVER Center at Caltech
David Goertson, led CAP-B10 and CAP-B22 at CLOVER
Xinhong Chen, led MaCPNS1 and MaCPNS2, contributed to M-CREATE at CLOVER
Miggy Chuapoco, led CAP-Mac at CLOVER
Hosted by Jason Nasse, Senior Scientist-Viral Communications at Addgene
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Research mentioned in the Q&A:
PHP.eB in Rat (Challis et al https://doi.org/10.1038/s41596-018-00...
Dayton et al 10.1038/s41434-018-0028-5)
The following AAV mentioned in the presentation are available at Addgene:
AAV-PHP.eB: https://www.addgene.org/103005/
AAV-PHP.VI: https://www.addgene.org/127847/
AAV-PHP.N: https://www.addgene.org/127851/
AAV-Cap.B10: https://www.addgene.org/175004/
AAV-F: https://www.addgene.org/166921/
AAV-PHP.B: https://www.addgene.org/103002/
AAV-PHP.S:https://www.addgene.org/103006/
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0:00 - Intro
2:10 - Main Presentation
20:18 - Q&A Introduction
20:57 - What advice would you give to someone just starting to use systemic AAVs in their experiments?
23:20 - Did Adenoviruses and AAV's co-evolve, where the same sort of utilization of diversity should therefore be possible with Adenoviral vectors?
24:18 - Is retro-orbital or tail vein more reproducible in your hands? we have had lack of consistency between users in both.
25:28 - Do you have any plans to optimize capsids to target microglia?
26:29 - You said AAV-CAP-B10 is better for Glutamatergic neurons, how about GABAergic neurons? Has anyone seen that?
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 LE 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, any 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 retrograde 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 AAV's?
43:55 - I was just wondering how we can 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 or leaky BBB influence the expression of PHP.eB? g.g., the circumventricular organs in the CNS
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
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