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  • Cluster analysis for hypothesis veryfication

    Hi,

    I am new in the field of cluster analysis of the RNAseq data and would like to ask more experienced guys if the strategy I plan to apply makes any sense.

    The hypothesis I want to verify states:
    The sorted cells from young knock-out (KO) mice have transcriptome profile resembling the cell from old wild-type (WT) mice.
    In other words, I suppose that KO cells show premature aging.

    Experiment scheme:
    I did RNAseq with 4 groups. 4 samples/group :
    1. WT - young
    2. WT - old
    3. KO - young
    4. KO - old
    Differential expression done by DEseq2.

    Now, does it make sense to use supervised sample-clustering to verify the hypothesis?
    I am thinking to select the "classifier genes" by comparing WT young and WT old - to select marker genes for aged phenotyped. I have around 1000 genes significantly changed by DESeq2.
    Then, based on this "classifier genes" I want to make supervised clustering (k-mean?) of all WT and KO samples, to see if young KO cell cluster together with old WT cells.
    Can I verify my hypothesis by this strategy? If yes, what tools (R packages or other programs) for supervised clustering do you recommend?

    I would be grateful for any help nad tips.

    Best,
    Krzysiek

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