Hi! I loved your poster it was very interesting! Is it possible to have multiple genes involved in DNA repair? if so, did you test any other genes or would you consider that process for future directions?
Thank you for your feedback! There are definitely many genes, and therefore proteins, that are involved in DNA repair as it is a highly regulated system. sRNAs are just one category of proteins that have been pinpointed for testing. There were several sRNA genes that our class as a whole performed this experiment on. For the future of my group’s findings, more in depth studies of what Rsp1 specifically does will be a reasonable area of focus for the future.
Hello! We did not do any western blotting for my group’s research. We did recommend that it be done in the future to confirm protein levels post DNA damage. As for what western blotting is, it is similar to gel electrophoresis of DNA in that it is a process that separates proteins by size. By looking at western blotting of the Rsp1 protein before and after damage, we would be able to confirm weather the increase in expression of mRNA goes all the way to the translated protein.
Hi Emma, T. thermophila is a good model organism for this experiment because of several reasons. First, it is a single celled eukaryotic organism which makes it affordable and easy to work with in the lab. Multicellular organisms are much more complicated to work with as I’m sure you can imagine. In addition, T. therm’s genome is highly conserved in higher eukaryotic organisms so as this research is expanded upon, it can potentially be applied to human DNA damage repair. Finally, T. therm has a naturally robust DNA repair pathway making it an excellent organism to use for this type of research question.
Hello, We used two different softwares for designing our primers called BLAST and Primer3Plus. You input the sequence of your gene of interest into these tools and they essentially look for areas where primers will potentially bind all while considering the risks of things like secondary structure formation. Once we chose our primers based on certain criteria, we ordered them to be made by a company that makes primers!
Hello, That is a very big question! To my knowledge, the Rsp1 protein would need to be expressed in a number of various conditions to observe the outcomes in cells with DNA damage with or without this protein present. In another study we read this semester, this was accomplished by using siRNAs that essentially chop up the mRNA for your specific protein before it can be translated. There are dozens of questions to answer in different contexts before stating a specific role of this protein but that would potentially be the next steps.
curesymposium.org 
Hi! I loved your poster it was very interesting! Is it possible to have multiple genes involved in DNA repair? if so, did you test any other genes or would you consider that process for future directions?
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Thank you for your feedback! There are definitely many genes, and therefore proteins, that are involved in DNA repair as it is a highly regulated system. sRNAs are just one category of proteins that have been pinpointed for testing. There were several sRNA genes that our class as a whole performed this experiment on. For the future of my group’s findings, more in depth studies of what Rsp1 specifically does will be a reasonable area of focus for the future.
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Great poster! I do not understand though what a western blot is or how it is performed?
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Hello! We did not do any western blotting for my group’s research. We did recommend that it be done in the future to confirm protein levels post DNA damage. As for what western blotting is, it is similar to gel electrophoresis of DNA in that it is a process that separates proteins by size. By looking at western blotting of the Rsp1 protein before and after damage, we would be able to confirm weather the increase in expression of mRNA goes all the way to the translated protein.
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Hello! Is there anything specific with the tetrahymena thermophila that made you choose it to perform this research on?
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Hi Emma, T. thermophila is a good model organism for this experiment because of several reasons. First, it is a single celled eukaryotic organism which makes it affordable and easy to work with in the lab. Multicellular organisms are much more complicated to work with as I’m sure you can imagine. In addition, T. therm’s genome is highly conserved in higher eukaryotic organisms so as this research is expanded upon, it can potentially be applied to human DNA damage repair. Finally, T. therm has a naturally robust DNA repair pathway making it an excellent organism to use for this type of research question.
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How exactly are primers/designed and physically made?
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Hello, We used two different softwares for designing our primers called BLAST and Primer3Plus. You input the sequence of your gene of interest into these tools and they essentially look for areas where primers will potentially bind all while considering the risks of things like secondary structure formation. Once we chose our primers based on certain criteria, we ordered them to be made by a company that makes primers!
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Nice job, this was super interesting. How would you determine RSP-1’s specific role in DDR experimentally?
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Hello, That is a very big question! To my knowledge, the Rsp1 protein would need to be expressed in a number of various conditions to observe the outcomes in cells with DNA damage with or without this protein present. In another study we read this semester, this was accomplished by using siRNAs that essentially chop up the mRNA for your specific protein before it can be translated. There are dozens of questions to answer in different contexts before stating a specific role of this protein but that would potentially be the next steps.
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