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1) You have transfected siRNA against transcription factor A into cells that normally express the transcription...

1) You have transfected siRNA against transcription factor A into cells that normally express the transcription factor. These cells also contain a luciferase reporter gene that is controlled an enhancer that has multiple binding sites for transcription factor A.
a. What would be the effect on reporter gene activity
b. How could you determine whether RNAi acts at the level of mRNA or translation in this case?

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a. The reporter gene activity will be reduced because the siRNA will target the transcription factor A and repress its function. So it will not be able to bind the enhancer regulating the reporter gene. So reduced reporter gene activity.

b. Whether RNAi acts at the RNA level or protein level can be determined by checking the levels of mRNA through a quantitative PCR or a semi-quantitative PCR. A western blot can be done by using an antibody against that protein A to check the protein level.

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