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Full ORF DNA CLone
Hello again,
Excuse mty ignorance if this is a stupid question:
If you have a genedit disease based in the absence of a protein, Does a custom Full ORF DNA clone by plasmid of the missed protein could avoid or fix the disease?
Comments
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...potentially. There's no guarantee that the protein will be expressed.
For simplicity I quote genecopoeia's faq: "A correct DNA sequence with a promoter does not guarantee you will get protein expressed. Many factors contribute to the success or failure of protein expression: 1) Did you use the right type of expression vector for your expression host system? For instance, our pReceiver-M01 vector is for mammalian cell expression and pReceiver-B01 is for bacterial expression. 2) If the protein is a membrane protein, does it need target sequence for proper localization? 3) Will the over-expressed protein be toxic to the host? 4) Will the protein be folded into its correct three-dimensional structure for proper function and/or immunological assays? 5) Have you checked the sequencing to see if you get an alternative splicing variant?" -
thanks for the reply,
I would also add, 6) Does the receptor works properly?.
Would it be right?
In case point 1,2,3,4 ( ufff scares ), and 5 could be solved, point 6 would be impossible to avoid? -
Your best bet would be to test in a cell culture before you move to self-experimentation.