over-expression the PEPCK-C enzyme in the Human skeletal muscle.

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sarmed
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over-expression the PEPCK-C enzyme in the Human skeletal muscle.

please read this first:
http://www.case.edu/med/biochemistry/highlight_mouse.html
 
ok how can we do the same thing in humans.....your suggestions???

g a
g a's picture
 Hi Sarmed

 Hi Sarmed

It's unethical to fiddle with human genome. The article mentions it clearly as well. However, there are ways to genetically engineer the human genome for other purposes such as stem cell transfection for many gene deletions etc. You can look into that for further details.

sarmed
sarmed's picture
  ok .... so I was wondering

  ok .... so I was wondering :

1) Viral transfection is the best type for in vivo transfection as far as effectivity so what's gonna happen for the letiviral or adeno-virus after transfection is done and the targeted gene has been knocked-out?

2) Which one of the other methods comes in the second place

3) what is  the best way  to active immunize against some gene ... better than risking knock it out while we can only make the immune syste  produce some antibodies against it????
 

earonesty
earonesty's picture
Rough overview:

Rough overview:

For about $5000, I could make a lentiviral vector that slapped in a copy of human PEPCK with an Actin promoter.   Would work fine.   Not a knocout... this would be a vector that induced a recombination event to "kock in" the target gene.   Probably there's a CRISPR way to do this that isn't "shotgun", but lentiviral methods are mature and there are companies that can simply "build a lentivirus for you".   Then I'd need about $50K to purchasde and house pre-aged JAX mice, treat half with the vector, keep them alive and monitor basic stuff (psych and physical fitness).   Then I'd like another $20k for RNA Seq so I can prove that a) the vector works and b) it's having the effects intended.   Finally, I'd like to look at the mice, post mortem, for signs of vector induced cancer.  

Total experiment would be, without accounting for the PI's time, $75K.   That's an off-the-cuff overview of how someone might go about it.