Hello Academicians and Nobel Winners:
I have a basic logical question regarding RNAseq, the holy basis of bioinformatics
I am studying disease-associated genes in CNS (Central Nervous System) and I have knockout out a gene in HEK293T cell (easy to manipulate, transfection,etc.) Then performed RNAseq to get some insights (really just for insights, give a kick).
The results is below, I do get significant down pathways and genes, but since then I am confused, how can i get hints from this? all in all, 293T is not a CNS cell line, but I do not want to abandon this results. Is it associated with transcription factors? Post-transcriptional regulations? or what else?
1 answer
Keep in mind that these overrepresentation analysis are primitive. All it says is that with a given probability more genes than expected by chance overlap with the annotated pathway. Also, many genes are redundant and unspecific, so despite that the term might contain "neuronal" the genes could just be bystanders and exist in many pathways.
That having said, HEK is obviously not a neuronal cell. What you maybe get are genes that are under the same molecular similar regulation and if you knockout a key regulator, then many change, and this is similar to what happens in neuronal cells. Still, don't overinterpret it. It's a cell line, in culture for decades, malignantly transformed, not neuronal, so a lot of confounders.
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