Reference no: EM132927504

Study carefully the paper entitled ‘Gemin5 Binds to the Survival Motor Neuron mRNA to Regulate SMN Expression'

1. What does the abbreviation snRNP stand for?

2. Explain briefly,
a) what is meant by "pre-mRNA splicing"
b) why this process is essential in eukaryotic cells but not prokaryotic cells.

3. Where in the human genome is the SMN protein encoded?

4. In the abbreviation PAR-CLIP, what does IP stand for?

5. Explain the use of Protein G on Dynabeads in PAR-CLIP.

6. Explain
a) the use of streptavidin in pulldown of biotinylated RNAs.

b) why is this interaction used so widely in biochemical techniques?

7. Explain why a secondary antibody is used for the Western blots.

8. a) In which two ways is RNA interference used to down-regulate specific RNA species in this paper?

b) Which of these two techniques is best for long-term down-regulation?

9. Explain
a) what is meant by "the Ct method" applied to realtime PCR.

b) why is the Ct for SMN mRNA likely to be significantly higher than the Ct for beta-actin mRNA?

10. Which of the authors' observations support the suggestion that Gemin5 binding requires
a) a particular base sequence,
b) a specific structural feature in the SMN RNA?

11. What is the effect of stable expression of Gemin5 shRNA
a) on the level of Gemin5 protein?
b) on the level of SMN protein?

c) How do the authors confirm that the effect on SMN protein levels is mediated by Gemin5 binding to the SMN mRNA 3'-UTR?