For my PhD I've been designing plasmids which replicate like a chromosome and can be maintained extrachromosomally (even replicated and delivered to the daughter cells upon proliferation) in the human cell. This completely avoids any integration issues, but it has only been tested in cell cultures.
That sounds awesome. Just make sure the artificial chromosomes won't get into germ cells, else the enhanced human isn't compatible to "old" / "original" humans anymore and interbreed won't be possible I assume.
Of course this method does not allow replacing genes, only adding new ones, but still has a lot of potential IMO.
RNA silencing? Then you can knock out the gene. And add the same gene with another codon usage / or another gene from yeast with the same function.
As to AAV itself, yes; you'd expect that a cell pre-infected with an AAV
at its target locus would be immune to re-"infection" with the same
locus-targeting AAV.
But as it is not really seqence dependend, there may be a chance it integrates close to the iother integrated virus DNA?
In case it were sequence specific you could include the targeted sequence in the virus, so it integrates a new integration site :D
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