A collaborator of mine has shown data where siRNA knockdown OF A TRANSCRIPT occurs as early as 8 hrs post-transfection. I cannot seem to find any similar data online. Has anyone out there performed similar time courses with similar findings? It may be target- or siRNA-specific, but I am curious to know whether anyone can comment on this point and steer me to a publication.
The real question I am asking is how quickly has anyone seen a target mRNA go down with siRNA transfection?
Emphasis should be more on the protein's half-life as RNAi stops de novo protein synthesis.
siRNA targets mRNA as soon as it reaches cytoplasm and interacts with the respective mRNA. However, the change at protein level differs from protein to protein depending on half-life or stability of protein. E.g. total p38 MAPK has relatively long half-life compared to L-type calcium channel, and therefore, after siRNA transfection it takes longer time to detect reduction in p38 protein compared to L-type calcium channel.
Was it a full KD? because 8h would be damn quick.
I got an almost full mRNA KD at 15h post-transfection...and I had found it very quick at the time. I had a second siRNA targeting the same transcript which was not that efficient and required more time to reach the same depletion level.
I had read something on Thermo Fischer's website where they said that as early as 4h post siRNA transfection they could observe a moderate but significant KD of the target gene, but full KD was attained between 24 to 48h.
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