Since muscle fiber nuclei aren't turned over in the same manner as
most other cells, due to the syncytium they form, the plasmid keeps
expressing the protein for quite a while, barring promoter methylation
or other forms of downregulation. In most other organs, transfected
cells will eventually die and be replaced, so you lose the plasmid
over time. I had considered using CpG depleted and/or muscle-specific
promoters, both of which might increase the duration of expression,
but the unbridled power of a wild-type CMV promoter has never been
exceeded. I can't say for sure how often/many these injections would
need to be, that's why I want to try it out.
As another exploitable characteristic of muscle syncytium, the
transfected nuclei spread along the entirety of the fiber's length,
and the protein will diffuse into the immediate area to sequester any
local myostatin, in a paracrine fashion. As there will likely be a
need for multiple injections per muscle group, those can be spread
laterally to reach most of the fibers.
I would be totally supportive of huffing the plasmids instead, but I
can guarantee that the expression levels will be extremely low. Also,
I'd like to avoid any off-target effects from follistatin, unlikely as
they may be.
> Yeah but how often would you have to inject/electroporate? I think
> most people would rather inhale a few milligrams of DNA aerosol
> through their nose every once in a while. It also seems like with
> electroporation of muscle you'd get uneven expression, and thus
> develop muscle deformities/disfigurement.
>
> --
> Nathan McCorkle
> Rochester Institute of Technology
> College of Science, Biotechnology/Bioinformatics
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