What the all the reads stand for?
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7.0 years ago
evolighting ▴ 20

If it tells us the RNA amount at the moment of cells being processed?

Or there are cells reaction to the processing also being captured?

Would all these things count?

RNA-Seq • 1.0k views
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7.0 years ago

It's a bit difficult to parse out what you're really asking, so I'll interpret it as, "What signal or state is standard RNAseq capturing? I this dynamic, steady-state, or something else?"

The answer is that you're mostly capturing steady state levels, though these can be effected by experimental treatment (e.g., knocking out a gene). While you can try to get a bit more dynamic, with things like RNAvelocity, that's generally not what RNAseq is telling you. If one wants to capture RNA dynamics, then typically things like GROseq are used to capture active transcription (or RiboSeq for translation). I presume someone has thought of a clever way of measuring RNA turn-over en mass too, but I'm not familiar with it.

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For metabolic RNA-Seq, you may have a look at Herzog et al SlamSeq. The RNA is labeled which leads to certain mismatches at the end. The amount of mismatches over time corresponds with the RNA's anabolic and catabolic dynamics.

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Cool, I was wondering if there were any new label-based methods, I foresee someone over hear wanting to do that in the next year or so.

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AFAIK, there is an Osmium-based method, but it interferes a lot with the cells' metabolism. In the linked paper, they show how to deal with their compounds' toxicity.

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