siRNA cell function examples - (Mar/11/2009 )
Can anyone suggest some good studies providing examples of study of gene function using siRNA in which the gene functions aren't already known?
First you need to get a rough idea of what the gene does by looking into its protein structure. If it contains a particular domain, then you know what the domain is for and what function of other proteins with the same domain have. Once you have an initial idea, you would know what phenotypic changes you are going to look for after RNAi knockdown of the gene such as cell cycle changes, apoptosis, etc.
Another way of screening for unknown genes that may be important for a particular biological process is RNAi library screening. For example, you want to know what genes may be important in regulating mitosis, you do a high throughput RNAi experiment (basically you knockdown every genes in the genome), and FACS analysis. If mitosis is changed in a sample, then you can trace back to find out which gene has been knocked down.