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Gene Therapy Vector systems(13 Votes)
Non-viral  15.38% -2 Votes
Retroviral  7.69% -1 Votes
Lentiviral  7.69% -1 Votes
Adenoviral  7.69% -1 Votes
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Herpes viral  23.08% -3 Votes
Other  7.69% -1 Votes

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Gene Therapy Vector systems

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parvoman

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Since this seems to be the forum that deals with gene therapy, I thought I'd see how many people are working in each of the main fields. I realise that the chances are that you will be using more than one approach, if so please select the one that you consider youself to be most expert in.

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 Posted Aug 03, 2005, 21:23 PM
COHscientist

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I am working on AAV system.
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Posted Feb 03, 2006, 21:23 PM
parvoman

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COHscientist said:
I am working on AAV system.


Are you examining wt AAV biology or using AAVs as transduction tools?

I used to work on AAV biology but have now switched to using AAVs (along with Lenti and Ad vectors) as transduction tools. Am interested in transducing macrophages with siRNA cassettes.
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Posted Feb 09, 2006, 18:27 PM
Tracy

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parvoman said:

I used to work on AAV biology but have now switched to using AAVs (along with Lenti and Ad vectors) as transduction tools. Am interested in transducing macrophages with siRNA cassettes.


We are now using AAV2 as a vehicle to introduce other viral genes into mouse, hoping to get better immune response.
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Posted Feb 11, 2006, 3:47 AM
parvoman

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Tracy- Are you using ss or ds rAAV? And which serotype(s)?
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Posted Mar 30, 2006, 14:02 PM
Tracy

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parvoman said:
Tracy- Are you using ss or ds rAAV? And which serotype(s)?


We are using ds rAAV2 in our lab and another lab.
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Posted Mar 30, 2006, 15:12 PM
parvoman

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Tracy said:
parvoman said:
Tracy- Are you using ss or ds rAAV? And which serotype(s)?


We are using ds rAAV2 in our lab and another lab.



Have you looked to see how early you can get peak expression with the ds AAVs? The reason I ask is that the macrophages that I might try to transduce only live for about 10 days post transduction. It would be good if expression were reasonable within the first 36 hours.

Thanks
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Posted Mar 31, 2006, 15:39 PM
Tracy

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We always harvest the virus after 72hours post-transfection. So I guess this is the peak time.
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Posted Mar 31, 2006, 21:11 PM
parvoman

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Sorry- I was refering to the infection you do using the rAAV in mice. How long after injection of rAAV have you been able to get good expression? Sorry about the confusion.
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Posted Apr 03, 2006, 19:40 PM
Tracy

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parvoman said:
Sorry- I was refering to the infection you do using the rAAV in mice. How long after injection of rAAV have you been able to get good expression? Sorry about the confusion.



In our case, we used rAAV for boost in the vaccine regimen. We did two boost one month apart and after 21 days, we collected spenocyte for CTL response. So we really haven't tried to test what you asked. But I remember others have done that, and seems after one week they could detect the expression (If I remember correctly).
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Posted Apr 04, 2006, 18:59 PM
scolix

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transgene expression using AAV-1 is much faster than the usual AAV-2.

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Posted Jun 16, 2006, 15:49 PM
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scolix said:
transgene expression using AAV-1 is much faster than the usual AAV-2.



Why is that?
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Posted Jun 16, 2006, 23:47 PM
scolix

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not sure abt the exact reason. Mayb AAV-1 is degraded slower than AAV-2.

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Posted Jun 19, 2006, 15:06 PM
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scolix said:
not sure abt the exact reason. Mayb AAV-1 is degraded slower than AAV-2.


Adeno-associated virus (AAV) serotype 1 (AAV1) has been shown to be more effective than the well-studied AAV serotype 2 (AAV2) in muscle gene transfer. Replacement of amino acids 350 to 430 of AAV2 VP1 with the corresponding amino acids from VP1 of AAV1 resulted in a hybrid vector, termed AAV-221-IV, which behaved similarly to AAV1 in vitro and in vivo in muscle.

http://www.liebertonline.com/doi/abs/10.1089/hum.2006.17.46
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Posted Jun 19, 2006, 23:04 PM
scolix

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We have seen better expression using AAV-1 in neuronal cultures and to an extent even invivo comapred to AAV-2
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Posted Jun 20, 2006, 13:57 PM
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