How to silence two different alternative splicing sequences - (Jan/25/2005 )
I need to silence 2 different alternative splicing sequences. does anyone knows how can i do it?
This is a common problem and one that traditional siRNA design algorithms cannot manage. Our siRNA design service can, however, manage this. THe way out siRNA design works is to actually map RNA for accessible regions, and then siRNAs are designed against these accessible regions. As the entire length of the RNA is mapped, accessible regions within those alternatively spliced exons could be picked, ensuring that specific knockdown of your specific isoform was achieved. PLease look at http://www.expresson.com for more information or give us a call on 44 131 527 4607 to discuss it further.
one technique developed by j. summerton could help you. He developped morpholinos which are antisenses nucleotides modified by addition of phosphoroamidate residue. I'm not a chemist so i can't spaeak a lot of this radical. But they work as blocking traduction in cellulo and are completly resistant to exonucleases. A bad side : their are a little expensive. You should get 400$ minimum for one
For more information you can go to the website of gene tools inc.
they have a great explanation of their product.
Send me an email if you want more.
In my opinion, the best way to ensure you are silencing the two alternative spliced mRNAs separately is to design your siRNA by eye. I have designed all of mine like this and can very specifically knock down alternatively spliced mRNAs.