Hi all,
A very quick question.
I have sorted human T cells from blood using anti-CD3E, CD4 and CD8a antibodies. We did single cell seq on those sorted cells.
Analysing the results, I do not see any expression of mRNA CD3E (neither D nor G).
I know there are discrepancies between mRNA and protein levels, but is this so dramatic?
Has anybody done purified T cells and had the same problem? Or is this a problem of my data/sample?
Thanks!!
Jaz
Hi Jazmina,
My immunology courses are far behind me, but if I remember well CDs are membranary immunoglobulins involved in cell-cell communication, CD standing for Class od Differenciation.
This means that these membrane proteins are markers for differenciated cells. I guess the mRNA coding for CD3 is transcribed during the differenciation, and once differenciated, CD3 proteins remain, while mRNA is degraded.
We have similar observations in plant cells, where cell wall modelling enzymes are transcribed in early differenciation phases - in the distal part of the root elongation zone for example - and once the amount of enzyme is sufficient, transcription stops, but enzymes still remain active and work to ensure cell expansion.
Cheers,
Oliv
I usually find that mRNA and protein levels do not corelate. Usually mRNA goes up and then 8-12 h after protein level peaks but mRNA level drops. This all depends on the protein. Histones are translated fairly quick so mRNA level is more similar to protein levels while SPRTN protein and its mRNA do not corelate. Also if protein is highly stable it will outlive its mRNA through the cell cycle
Jaz,
differences in mRNA and protein levels are obviously normal and more pronounced for proteins with a reduced turnover/ half life.
We have done some experiments using specific hormones and other substances with mRNA time lines and you would be suprized to see what happens at different time points compared to the normal 24h experiments.
However, for your specific observation, you might wanna find out half life of your protein and check this article:
Article Genome-wide analysis of mRNA decay in resting and activated ...
and the supplements.
Best
Boris
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA