shRNA cloning - (Jun/22/2006 )

hello every one,
i have to express siRNA in plants. can you please suggest me what vectors to use for siRNA cloning and better expression in plants.iam new to this field.among dsRNA or siRNA which one will be effective?i will be thankful to you if you suggest how to design siRNA also.I have gone through the rules on how to design siRNAs.
Restriction site+21sense RNA+9nt spacer+21antisense RNA for one oligo and the same for another oligo in reverse order.is it ok.
awaiting for some help,
thank you in advance
cnu

There are a couple very good resources online. One is the Genscript company website, the other is the Open Biosystems company website. They have different approaches to the design of hairpins, although I can't suggest one method being better than the other. Here you can find expression vectors and tools to help design your hairpins.

Invitrogen site helps u to design siRNA or shRNA. There r also many sites from companies which help u to design ur shRNA.

Your idea of shRNA design seems ok" Restriction site+21sense RNA+9nt spacer+21antisense RNA for one oligo and the same for another oligo in reverse order".

Some people use 7 or even 5 bp as spacer. But i guess it shouldne b a problem.