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Designing new RNAi


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#1 july

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Posted 24 June 2009 - 10:43 PM

Hello,

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 :D

#2 pcrman

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Posted 25 June 2009 - 06:21 PM

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.

#3 july

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Posted 25 June 2009 - 11:27 PM

Thank you for the answer.
You are right.
Maybe there is something that I just cant understand. :huh:
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.

Thank you




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