Recently we overexpressed a targeted gene on the lentivirus vector. However, through repeated analysing, the elevation was validated on protein level by WB while not on mRNA level by qRT-PCR. Interestingly, sometimes the mRNA expression downregulated 30% and we wonder how it happened？
Hi Hong
probably a problem of techniques before digging in the science explanations.
dis you test your primers by in silico PCR?
you can test the UCSC tool (among others) with the right genome and target (UCSC genes will allow you an "amplification" on mRNA): http://genome.ucsc.edu/cgi-bin/hgPcr
all the best
fred
Hi Frederic,
we have 3-pair primers for qPCR experiment. And all these results showed the same.
you need to test them all therefore to be sure the target is the one you're expecting.
fred
The mRNA abundance of a specific gene may not have a linear relationship with the expression level of the translated protein. Factors such as mRNA degradation, protein degradation, and protein modification may cause mRNA expression levels to be inconsistent with the protein abundance. In addition, the expression levels of many genes change over time.
Vogel and Marcotte (2012) have a very good review of this issue.
http://www.ncbi.nlm.nih.gov/pubmed/22411467
Check it out Thanks
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