9 thoughts on “C27 – Pack

    1. Excellent question! It was found that my gene hyproc14 doesn’t actually have any human or mouse homologs. So, If I had been able to confirm hyproc14’s role in double strand break repair in DNA, we could use gene knock-in with mouse models to test efficacy of this gene as a contributor to DNA damage repair in mice. Subsequently following success, this could be continued in clinical trials with humans with cancer or other illnesses that cause excessive amounts of double strand breaks in DNA.

  1. This was a good presentation. Can you explain a little bit more about using gene knock-outs and knock-ins to identify the function of hyproc-14?

    1. Thank you, and absolutely! The way that we tested for these untested genes in DNA damage repair is by inducing double strand breaks in DNA from Tetrahymena and checking for an increase in gene expression which could be seen using Rad51 as the positive control. Using a different approach, we could use gene knock-out of the hyproc14 gene. Then, we could test the cDNA from the DNA that lacks hyproc14 as well as DNA that still contains the hyproc14 gene. We could then compare gene expression of Rad51 again between these two groups. If gene expression of Rad51 was higher in the subset of DNA that lacked hyproc14, it could be concluded that hyproc14 is involved in DNA damage repair, but due to being knocked out, Rad51 expression had to increase even more to compensate for the absence of repair that hyproc14 would have accomplished. With gene knock-in, we could simply duplicate this exact experiment using mouse cDNA with the hyproc14 gene knocked-in using vector DNA.

  2. Amazing presentation! Can you explain how gene-knock out would be a suggested further direction to take? You explain gene knock-in, but I am a little confused on the gene knock-out mechanism.

    1. Thank you! Someone else had the same question. So to summarize, we could have two subsets of cDNA from Tetrahymena. One subset as the control that still has the hyproc14 gene. The other subset that has the hyproc14 gene removed through gene knock-out. Next, we induce double strand breaks into both subsets of cDNA using hydroxyurea like we did in this experiment. Then, we test for Rad51 expression in both subsets. If, hyproc14 is involved in double strand break repair, gene expression of the cDNA lacking hyproc14 should be greater than the control group. If hyproc14 is removed and unable to perform DNA damage repair, Rad51 levels may increase to compensate.

    1. Great question Tasnim! This is true for all of the genes researched in our lab, but the main thing is that hyproc14 expression spikes during conjugation of Tetrahymena. Conjugation is the transfer of genetic information between two bacterial cells and conjugation is known to induce the SOS response in DNA damage repair. As a reference, Rad51 expression also spikes during conjugation and Rad51 is a known repair factor of double strand breaks in DNA.

  3. How can hyproc14 being involved in double strand break repair be beneficial in medicinal field?

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