I just want to know about the necessity of the MTT assay for cell viability, even if no abnormal observations were detected with the modified siRNAs.
Generally, there is no requirement for MTT assay. However, if your particular experiment is with some type of especially dangerous siRNA or something, then the situation may be different.
Modified siRNA serves the purpose to achieve high stability, efficiency, and effectiveness to target the GOI. The incorporation of such modifications in siRNAs might then give rise to molecules with enhanced cytotoxicity. So, it will be better if you perform any cytotoxicity assay.
Even, I feel somewhat in /overexpression/silencing/knockdown studies, transfection reagent also causes unnecessary effect on cell viability. So, better to do any cytotoxicity assay.
Of course you should focus on expression and targets. However, siRNA can affect cell viability, even the transfection itself can do so (as mentioned above), so you'd need to test cell viability.
The question could be more understandable if you clarify what do you mean by (no abnormal observations were detected)
Iman Hassan Ibrahim
Sir,
Thank you for your suggestion. By "no abnormal observations were detected", I mean that the RNAi activity did not alter drastically. As well as, when cell uptake assay was done, the cellular internalization efficiency was also similar to the native siRNA. Hence, I used the word as no abnormal observations were detected.
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