Designing new RNAi - (Jun/25/2009 )
I am new here.
I dont understand how can I Design a new RNAi, that will silence the genes that I choose.
Another question lets say that I want to silence three different gens.
I need to use three different RNA sequences that are complementary to the specifics genes that I want to silence right?
What to I add between the RNA sequences? I need something so the Dicer will know to cut there right?
I hope I am not to far off the answer.
Thank's for the help
Before you start to use RNAi in your project, you really need to get a basic understanding of how RNAi works, then you will be able to ask some specific questions we can answer here.
Thank you for the answer.
You are right.
Maybe there is something that I just cant understand.
I am really new in all the lab work (starting now my M.sc) so any help will be very appreciated.
If you could check if what I am writing is correct it will be very helpful.
As I understand it:
There is a dsRNA that enters the cell.
The Dicer cuts the dsRNA to SiRNA and MiRNA.
A RISC SiRNA complex created.
The RISC complex unwind the SiRNA so the uncomplimentary strand leaves the complex.
The ssRNA (the guide strand) he guides the endonuclease ( the Argonaute protein) that inside the RISC complex.
The Argonaute will cut the mRNA that are homology to the sequence of the ssRNA.