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-   -   What is RNA silencing..???????????? Wrt to RNA interference? (http://www.molecularstation.com/forum/rnai-sirna-forum/87740-what-rna-silencing-wrt-rna-interference.html)

Neel 03-24-2013 05:34 AM

What is RNA silencing..???????????? Wrt to RNA interference?
 
Can anyone Explain RNAi in simpler term????

luisillo 03-25-2013 05:02 PM

Re: What is RNA silencing..???????????? Wrt to RNA interference?
 
You have a peptide, say WT1, that is being expressed a lot in a given cell line, but you want to silence the expression of that peptide. Once you know the mRNA sequence of WT1, you order an antisense oligonucleotide, complementary to a part of WT1's mRNA sequence. Next, you introduce that oligo in your cell line by the method you see fit (eletroporation, etc.).

Once in the cytoplasm of the cells of interest the oligo will bind to WT1 mRNA, a part of this will become double stranded impeding its further translation. That will silence the expression of WT1 as long as there are antisense oligo in that cell's cytoplasm. That is RNA interference.

Jon Moulton 03-29-2013 04:09 PM

Re: What is RNA silencing..???????????? Wrt to RNA interference?
 
Lusillo, your description sounds like a description of steric-blocking antisense. RNAi involves the activity of an Argonaute-family protein to silence gene expression; short dsRNA loads onto Argonaute, one strand is cleaved and falls away and the other strand aids in recognition of sites on RNA triggering translation inhibition or cleavage of the target RNA. Your description more closely matches the activity of steric-blocking antisense like 2'-O-methyl phosphorothioate RNA, locked nucleic acids, peptide nucleic acids or Morpholino oligos.

Summerton J. Morpholino Antisense Oligomers: The Case for an RNase-H Independent Structural Type. Biochimica et Biophysica Acta 1999 1489: 141-158

luisillo 04-01-2013 05:25 PM

Re: What is RNA silencing..???????????? Wrt to RNA interference?
 
Yes, I described an antisense activity. Guess I got mixed. Thanks Jon for correcting me and for the info and the reference.


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