Following is a scientist's responses to the question, "If an RNAi experiment results in insufficient gene knockdown, do you try the experiment again? If so, what approach do you use to improve the knockdown?"
For a complete list of scientists' responses, read the March issue of Genome Technology, a GenomeWeb News sister publication.
"If the RNAi sequence is getting into the cell and just not knocking it down enough, we will try another sequence or a combination of sequences. Invitrogen sells 'smart pools' of RNAi's consisting of four or five sequences. Sometimes you can get better knockdown just by shifting the sequence a couple of nucleotides. If the RNAi isn't getting into the cell (test by uptake of a labeled oligo, e.g. Block it by Invitrogen) we will try other transfection reagents or may engineer into a viral vector (e.g. lentivirus, herpesvirus, etc.)."
Johnson & Johnson