1)You perform an SiRNA screen to understand what components of the recombination
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Question
1)You perform an SiRNA screen to understand what components of the recombination pathway might be found. Draw out an experimental approach to finding these components. Make sure to label your reporter, note its key features and how it functions to report recombination.
2)You get the following data
Recombination Rate Wild Type: 20%
Recombination Rate Candidate gene siRNA: 50%
Is your gene a suppresssor or a mediator?
3)Name at least three proteins that you would expect to find in the experiment
4)You think that recombination might be affecting the overall conformation of the chromosome. Describe the method you would use to confirm this hypothesis. Bonus: Describe another method to confirm your results.
Explanation / Answer
Various components of recombinant pathways are:
Experimental approach for SiRNA screen is as follows:
2. Data as provided:
- Recombination Rate Wild Type: 20%
- Recombination Rate Candidate gene siRNA: 50%
Here siRNA is 50% so this will definitely be a suppressor gene as siRNA also includes co-suppression by silencing.
3. Three proteins that are probably found in the silencing experiment are:
4. As per the process involved in siRNA silencing mechanism, I believe the siRNA also affects overall conformation of chromosome as the main causer will be the dsRNA cleavage using the Dicer protein which mainly cleaves the RNA into fragments and ultimately affecting the transcriptional activities (protein synthesis). Hence the silencing mechanism is also affecting the conformation of chromosome, as many viral infections to which eukaryotes were immune earlier are known prone hence there is regular evolution (conformational changes of chromosomes are happening) of chromosome.
Other methods include High throughput screening which involves loss of function screening.
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