Designing siRNA expression vectors - (Aug/25/2005 )

Hey,

I'm trying to design a adeno 1.0-CMV based siRNA expression vector.However, I'm not being able to provide the RefSeq Accession number of the cDNA sequence(has to be like NM_012345 or XM_012345)in turn I only have a accession number.In result, I could find a possible siRNA target region in the database. I don't know whether it's because the number I input is wrong or the database doen't include it at all.

Also, I heard from other people that u always design several(2-4) target region for silencing to make sure one of them would get a 75-95%silencing.Thus is there any special consideration in choosing the area or just randomly pick one or two?

Great appreciation

Plasmid

>>In result, I could find a possible siRNA target region in the database.

What database? Do you mean "I could not find ..."

>>I heard from other people that u always design several(2-4) target region for silencing to make sure one of them would get a 75-95%silencing.Thus is there any special consideration in choosing the area or just randomly pick one or two?

Yes, people usually design at least two siRNAs for each gene just in case one doesn't work. There are no particular rules regarding which area along the coding sequence to pick siRNA targets and any region should work, but I think it is better to stick to exons (not UTRs).

Thank you very much!