User:
Catherine Taylor, University of Waterloo, Canada
Project:
Using siRNA knockdown to identify proteins involved in apoptosis
Problem:
As in Ruiz-Vela's case above, SiRNAs can induce off-target effects.
Controls:
Taylor always performs separate experiments with different siRNA sequences - each designed to knock down the same mRNA, but by targeting a different part of the transcript. It's "absolutely crucial," says Taylor. "I never do any experiments without it." She learned her lesson when she once used just one siRNA to suppress a gene and obtained a result she was expecting from previous work. "I was convinced it was correct, and I didn't include the second siRNA right away. When I did, I found it didn't behave the same as the first siRNA," she says. "Sure enough, it was due to some other effect."
Taylor also uses one of these triplets for a rescue experiment - the ultimate siRNA control. She ...
