How does identifying the proteins involved in DNA damage prevention and repair pathways contribute to drug development for diseases related to DNA damage/what role does the protein play in DNA repair? Great presentation!
We were more looking at the process of DNA damage prevention or repair as a whole to gain more overall insight, we weren’t looking for a direct medical use but the hope is that by learning more about the process and what is involved maybe we could find a medical application. A possible connection that I have no real basis for could be if there is a similar protein to Twi7 in humans and we understood the pathway then maybe we would see an increase in cancer in people where it is not present if Twi7 is involved in DNA damage/repair, then maybe we could find something that could replace it maybe a drug or something to help prevent cancer. At this point we are not sure what the protein does in the pathway that is what we are trying to study, this suggested that Twi7 might not be involved in DNA double stranded break repair.
So in the PCR that we used the thermocycler is run for a specific set number of cycles which will give a different number of copies of the desired output based on the initial concentration, now different concentrations of the sample will show up well on a gel at different numbers of cycles. we could run many PCRs to test a lot of different number of cycles but it is easier to use qPCR in this case because it measures the amount of DNA product after each cycle. this means it could tell us if the reason we didn’t see any expression of Twi7 before or after DNA damage was because maybe the level of Twi7 in the cell is very low and simply wasn’t enough to show up in our gel after the number of cycles we used. If it still didn’t show a change in expression this could also be useful in confirming the data we found that Twi7 does not seem to be involved in the repair of DSBs.
What would be the process to research wether a gene plays a role in preventing DNA damage? And how could you measure that? How does this connect to cancer research?
You know I am not really sure, I think that with this research combined with DR. Suzanne Lee’s research which showed an accumulation of DNA damage when Twi7 was disabled, could suggest that Twi7 is involved in the prevention of DNA damage rather than the repair. I am not sure if this would work but maybe we could also do a coimmunoprecipitation assay to see if Twi7 binds to some proteins which are known to be involved in DNA damage prevention, if they did coimmunoprecipitate this could tell us that they are involved in a prevention mechanism.
Great job! How would you test if Twi7 does single strand break repair, and what results with this future direction would remain consistent with your hypothesis?
To test if Twi7 might be involved in single stranded break repair we could repeat this study using an agent other that Hydroxyurea which causes single stranded breaks, an example would be H2O2 hydrogen peroxide which is known to cause single stranded breaks. Then we could repeat the PCR and gel and see if there is an increase of expression of Twi7 with the use of H2O2. If there was not an increase this could indicate that Twi7 is more likely involved in the prevention of DNA damage, or that it is just involved in DNA repair under different circumstances than either H2O2 or Hydroxyurea.
curesymposium.org 
How does identifying the proteins involved in DNA damage prevention and repair pathways contribute to drug development for diseases related to DNA damage/what role does the protein play in DNA repair? Great presentation!
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We were more looking at the process of DNA damage prevention or repair as a whole to gain more overall insight, we weren’t looking for a direct medical use but the hope is that by learning more about the process and what is involved maybe we could find a medical application. A possible connection that I have no real basis for could be if there is a similar protein to Twi7 in humans and we understood the pathway then maybe we would see an increase in cancer in people where it is not present if Twi7 is involved in DNA damage/repair, then maybe we could find something that could replace it maybe a drug or something to help prevent cancer. At this point we are not sure what the protein does in the pathway that is what we are trying to study, this suggested that Twi7 might not be involved in DNA double stranded break repair.
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What is the difference between qPCR and the PCR that you have previously used?
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So in the PCR that we used the thermocycler is run for a specific set number of cycles which will give a different number of copies of the desired output based on the initial concentration, now different concentrations of the sample will show up well on a gel at different numbers of cycles. we could run many PCRs to test a lot of different number of cycles but it is easier to use qPCR in this case because it measures the amount of DNA product after each cycle. this means it could tell us if the reason we didn’t see any expression of Twi7 before or after DNA damage was because maybe the level of Twi7 in the cell is very low and simply wasn’t enough to show up in our gel after the number of cycles we used. If it still didn’t show a change in expression this could also be useful in confirming the data we found that Twi7 does not seem to be involved in the repair of DSBs.
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What would be the process to research wether a gene plays a role in preventing DNA damage? And how could you measure that? How does this connect to cancer research?
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You know I am not really sure, I think that with this research combined with DR. Suzanne Lee’s research which showed an accumulation of DNA damage when Twi7 was disabled, could suggest that Twi7 is involved in the prevention of DNA damage rather than the repair. I am not sure if this would work but maybe we could also do a coimmunoprecipitation assay to see if Twi7 binds to some proteins which are known to be involved in DNA damage prevention, if they did coimmunoprecipitate this could tell us that they are involved in a prevention mechanism.
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What in particular does the qPCR do that the PCR you used wouldn’t? And how would this more focused information help your research going forward?
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Great job! How would you test if Twi7 does single strand break repair, and what results with this future direction would remain consistent with your hypothesis?
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To test if Twi7 might be involved in single stranded break repair we could repeat this study using an agent other that Hydroxyurea which causes single stranded breaks, an example would be H2O2 hydrogen peroxide which is known to cause single stranded breaks. Then we could repeat the PCR and gel and see if there is an increase of expression of Twi7 with the use of H2O2. If there was not an increase this could indicate that Twi7 is more likely involved in the prevention of DNA damage, or that it is just involved in DNA repair under different circumstances than either H2O2 or Hydroxyurea.
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