Suggestion for approaching multiple conditions scRNA-seq
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20 months ago
Eisuan ▴ 20

Hello everyone, I have a scRNA-seq dataset with control, condition 1 and condition 1 + 2 (sh). Condition 1 induces a phenotype that has been previously described and I was able to characterize it by adding some details given the higher number of cells in our dataset compared to previous works.

Condition 2 is expected to influence the "normal" phenotypes induced by condition 1.

I was suggested to perform an exploratory evaluation of the clusters I can recognize in condition 1 and in condition 2 alone. Unfortunately, I am not able to recognize dramatic changes. I know that the system that induces condition 2 works since differences can be detected in bulk seq.

How should I approach this analysis? I have seen approaches such as pseudo-bulk analysis, but I fear I have too few samples for having a robust estimation of dispersion (2 for each sample).

Thank you

scRNA-seq • 734 views
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Do you have 2 biological or technical replicates for each condition? Also, you mentioned you didn't find dramatic changes with your analysis. What did you do for that analysis?

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Hi! Two biological replicates. The previous analysis has been performed on Seurat.

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"performed on Seurat" is like saying that the repair was done with a hammer and skrewdriver. Seurat has plenty of functions, you need to be a bit more specific.

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Hi, I analyzed the datasets separately as follows:

  • control+condition 1
  • control + condition 1+2 (not all the three merged!)

I applied the standard worklow looking for markers using WSRT (find all markers).

The first dataset has been extensively characterized, I also compared DEGs between subpopulations and performed ORA and GSEA.

In the second I specifically searched for a subpopulation that I expected to be deregulated by condition 2. However, I still can find its markers.

I was wondering if there are more appropriate solutions to check if there are quantitative differences.

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