I do not have any experience working with LV and AAV's, but planning to use it for kd and overexpression of gene. I was wondering if LV and AAV's can be used as a quick alternative for Cre-ER for expression or kd of gene in tissue specific manner?
Also, if anyone has any information about how long does the AAV based expression persist in post-mitotic cells?
Thank you!
A while back I started similar work using lenti virus and the preliminary results were good, make sure you titrate your virus quantity to avoid toxicity. The plus side of lentivirus is the establishment of stable rather than transient transfectants
AAV lasts a long time in post-mitotic cells. Dogs that were treated with AAV still express FIX after >10 years.
I was using lentiviral mediated gene delivery during my PhD... These can be used for tissue specific under/over expression of genes, you have to engineer the carrier plasmid cautiously...Good thing about LV/AAV is you can use them for postmitotic cells too which cant be done for retroviral system. As Jonathan says AAV shows expression after 10 years that is something amazinng, I didnt know that, I knew it works for 2-6 months...So I think we can use it for tissue specific and long term expression ....
aavs-carried genes are expressed for very long term. In my experience, > 2 months, which is usually sufficient for experimental approaches. Specificity can be reached to a reasonable exent by combining the right aav serotype with a suitable mini-promoter with tissue or cell-restricted expression. not all minipromoters are "air-tight" but there a number that works pretty well. knock-down is also feasible with aavs, but the extent may be variable and never reaches 100%.
AAV-mediated expression persistence sometimes depends on AAV serotype and the promoter used, but I have seen AAV-2 GFP expression in rat retina after more than 1 year. A tissue-specific promoter will help with specificity but you might not get 100% targeting. Knockdown by siRNA/shRNA is well described using LV or AAV, but the effect wouldn't be exactly the same. KD is usually not 100% efficient with any vector and it is a very different mechanism than Cre-lox.
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