RNAi - possibilities and applications - (Mar/04/2008 )
So what can you basically do with RNAi? I mean knockout or "silence" the gene of interest and then what? You look for morphological changes in the cell? Quantify mRNA levels too see what changed if the knockout is viable? I mean what do people usually do with this technique? What are the possibilities?
-gorkin-
QUOTE (gorkin @ Mar 4 2008, 03:03 PM)
So what can you basically do with RNAi? I mean knockout or "silence" the gene of interest and then what? You look for morphological changes in the cell? Quantify mRNA levels too see what changed if the knockout is viable? I mean what do people usually do with this technique? What are the possibilities?
Cell proliferation, apoptosis, differentiation, morphology, cell adhesion, invasion, motility...the list is endless. You just need to think about your hypothesis and pick the right assay to use.
-miRNA man-
QUOTE (miRNA man @ Mar 5 2008, 10:55 AM)
QUOTE (gorkin @ Mar 4 2008, 03:03 PM)
So what can you basically do with RNAi? I mean knockout or "silence" the gene of interest and then what? You look for morphological changes in the cell? Quantify mRNA levels too see what changed if the knockout is viable? I mean what do people usually do with this technique? What are the possibilities?
Cell proliferation, apoptosis, differentiation, morphology, cell adhesion, invasion, motility...the list is endless. You just need to think about your hypothesis and pick the right assay to use.
Oh sorry, I meant with regard to certain metabolic enzymes.
-gorkin-
QUOTE (gorkin @ Mar 6 2008, 03:44 AM)
Oh sorry, I meant with regard to certain metabolic enzymes.
That narrows it down quite a bit. Assay options include testing the concentration of target mRNA, assaying the target mRNA's protein product (if you have an antibody), or measuring enzymatic activity of the target mRNA's protein product. The challenge is the hunt for off-target gene modulation, either as changes in off-target mRNA concentrations or as the more widespread translation inhibition of off-target mRNA; the former can be assessed by nucleic acid microarray, the latter is more difficult to measure.
-Jon Moulton-