Given the final level of Twi8 is low, and the literature indicated this would be the case, what was the intended goal when you picked this gene? What roles does this gene have that make it relevant to study in the context of DNA damage repair?
Dr. Lee provided a consensus gel that gave a bit of insight or “literature” that supported our results. We still did research to test change of gene expression in Twi8 given that this literature was also done by students and we were given an opportunity to add onto their findings. We researched sRNAs as sRNAS and their interacting proteins have been found to stabilize the genome and we have since understood that sRNA molecules are a powerful tool used by animals and plants for genome stability and our being researched for their role in the DNA repair pathway.
Hi Jack, great question. The primers annealed particularly well given that we did end up visualizing bands within our gene of interest to the different environment it was exposed to. In terms of actually making sure that the primers annealed correctly we followed a criteria using Primer3Plus that minimized our chances of creating different dimers or hairpins.
What implications could the finding that Twi8 gene expression remains unaffected after DNA damage in Tetrahymena thermophila have for future studies on DNA repair pathways?
Great question, while the Twi8 gene expression remains unaffected after DNA damage in Tetrahymena, researchers can find other possible uses of the Twi8 gene outside of the DNA damage repair pathway such as possible implications in genome stability or homeostasis. While this sRNA produced a finding that did not support our hypothesis, other students have found that their specific sRNA could in fact be involved in the DNA damage repair pathway!
Hi Joshua, great question. Tetrahymena Thermophila makes it a good model to study gene expression it is known to have interacting small RNAs that contribute to a number of functions within its cells such as self-splicing or catalysis of sequence-specific cleavage. At the same time Tetrahymena Thermophila has easily seen experimental advantages where they are easy and inexpensive to culture, and are amenable to biochemistry, microscopy, bioinformatics, and reverse genetics. Using this organism provided an opportunity to explore a broadly conserved and evolutionarily innovative biology.
Why do you think that the Twi8 gene not being expressed (or low) via PCR is the case?? If the literature indicates that this level is low what could possibly be done to see more prominent results?
Hi Kate, great question. Our lab group deduced that the Twi8 gene was expressed in a low quantity due to the dimness of the band from gel electrophoresis, and given that the literature indicates a correlation, it may be worth exploring Twi8’s involvement in the DNA repair pathway using qPCR or Northwestern blotting.
curesymposium.org 
Given the final level of Twi8 is low, and the literature indicated this would be the case, what was the intended goal when you picked this gene? What roles does this gene have that make it relevant to study in the context of DNA damage repair?
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Dr. Lee provided a consensus gel that gave a bit of insight or “literature” that supported our results. We still did research to test change of gene expression in Twi8 given that this literature was also done by students and we were given an opportunity to add onto their findings. We researched sRNAs as sRNAS and their interacting proteins have been found to stabilize the genome and we have since understood that sRNA molecules are a powerful tool used by animals and plants for genome stability and our being researched for their role in the DNA repair pathway.
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What other genes could you study in relation to Twi8 in the future to determine it’s role in dna repair?
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Why does those primers anneal particularly well to the chosen gene?
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Hi Jack, great question. The primers annealed particularly well given that we did end up visualizing bands within our gene of interest to the different environment it was exposed to. In terms of actually making sure that the primers annealed correctly we followed a criteria using Primer3Plus that minimized our chances of creating different dimers or hairpins.
LikeLike
What implications could the finding that Twi8 gene expression remains unaffected after DNA damage in Tetrahymena thermophila have for future studies on DNA repair pathways?
LikeLike
Great question, while the Twi8 gene expression remains unaffected after DNA damage in Tetrahymena, researchers can find other possible uses of the Twi8 gene outside of the DNA damage repair pathway such as possible implications in genome stability or homeostasis. While this sRNA produced a finding that did not support our hypothesis, other students have found that their specific sRNA could in fact be involved in the DNA damage repair pathway!
LikeLike
What about Tetrahymena thermophila makes it a good model to study gene expression of Twi8?
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Hi Joshua, great question. Tetrahymena Thermophila makes it a good model to study gene expression it is known to have interacting small RNAs that contribute to a number of functions within its cells such as self-splicing or catalysis of sequence-specific cleavage. At the same time Tetrahymena Thermophila has easily seen experimental advantages where they are easy and inexpensive to culture, and are amenable to biochemistry, microscopy, bioinformatics, and reverse genetics. Using this organism provided an opportunity to explore a broadly conserved and evolutionarily innovative biology.
LikeLike
Why do you think that the Twi8 gene not being expressed (or low) via PCR is the case?? If the literature indicates that this level is low what could possibly be done to see more prominent results?
LikeLike
Hi Kate, great question. Our lab group deduced that the Twi8 gene was expressed in a low quantity due to the dimness of the band from gel electrophoresis, and given that the literature indicates a correlation, it may be worth exploring Twi8’s involvement in the DNA repair pathway using qPCR or Northwestern blotting.
LikeLike