Systemic Administration of AAV PHP.S for Delivery to DRG

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Has anyone used AAV-PHP.S in neonatal mice (P1) via the intraperitoneal route for delivery to DRG?
In the original paper, they do IV admin and the transduction is great, but IP may be easier and it saves us some time while the animals come of age.

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@liz @tberta @thicunha

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@achamess we are producing these AAV in-hourse. We have performed the first experiment just to establish. @tberta is using AAV9 injected in the paw of neonatal mice and he saw transduction in the DRGs.

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We’ve tried retro-orbital dosing with a couple PHP.S viruses and have never seen transduction efficiency close to what was reported in the original paper. Typically only ~15-25% of neurons express the reporter in our hands, even with high titers.

I’ve been meaning to try intrathecal but it seems like that also has a low efficiency based on some published studies. Neonatal i.p. Is an interesting thought…I wonder if it would be any better than AAV9 though.

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Thanks a lot @james_maks for the info. I’m sorry it’s not working as expected. What promoter are you using? CAG? And are these in house preps or Addgene?

It appear efficacy is strain dependent too (in PHP.B), with C57 being the best

https://www.cell.com/molecular-therapy-family/methods/fulltext/S2329-0501(20)30215-1

https://www.cell.com/molecular-therapy-family/molecular-therapy/fulltext/S1525-0016(18)30025-X?{%24trackingTag}=&amp%3Belsca2=email&amp%3Belsca3=1525-0016_20180307_26_3_&amp%3Belsca4=

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@achamess We’ve tried with CAG, CBh, and both Cre-dependent and constitutively expressed constructs and basically get the same results.

We’ve run into problems with strain concerns using PHP.eB but with PHP.S, all mice were C57s or the transgenics were backcrossed and genotyped to be C57 congenics. In our hands, the PHP.S just doesn’t work that well dosed r.o.

We source our AAVs from VectorBuilder so that might be an issue, but PHP.eB/AAV9/etc. all work as expected.

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Hi @achamess,

I am planning to use AAV-PHP.S and/or AAV-MaCPNS1/2 for transducing DRG neurons. Have you tried IP admin at P1? How was the transduction efficiency? There are papers showing the AAV transduction is dependent on administration routes and mouse strains. I was wondering which injection method will work best for me (IP, retro-orbital, or tail vein injection)? Have you tried deliver AAVs during P21-P28? Thank you!

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Let us know hot goes. I haven’t tried. I did some retrograde from the paw in adult mice and it was underwhelming. I think you have to just give it a try. Make sure your virus is prepared by a reputable source. I think this make a big difference.

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I will let you know how it goes. Thank you for the info!

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