To knock out a gene would basically mean to make it inactive or remove the gene itself so that no proteins can be synthesized from it. This allows for researchers to see how functions in the cell change without the presence of the gene in question. One way this is done is by using CRISPR-Cas9, but I am unsure of the method that was used when Dr.Lee found that the knockout of Twi7 leads to no DNA damage accumulation.
You mentioned in your methods that you were creating gene specific primers, and that sounds really cool. How did you do that? If you have time to fit it in, you should what methods you used to make gene specific primers.
Yes, it actually is quite a fascinating process. In order to design our primers, we used the website called primer3plus, which allowed us to find a set of primers that would work best for the gene we have been given. And then, we used BLAST which allowed us to evaluate if the primers we chose would be found in other Tetrahymena genes. When the results showed us that they would not, and the primers hit the qualifications that they needed to, we were able to order them and they were delivered to the lab.
Many cancer therapies used today are targeting cancer cells to cause DNA damage and therefore kill the cell and make it nonfunctional. With our results, we know that the Twi7 gene likely does not play a role in the DNA damage response pathway, meaning it does not aid the cells in fixing the damaged DNA. It is possible that researchers could knock out the other Twi proteins that were found to play a role in the response pathway, but keep Twi7 and see how the cell responds. This could lead to a better understanding of how to target and kill cancer cells. In terms of future research that could be conducted based on the results that we got, studying the role of Twi7 in other parts of the cell would be the most beneficial.
When looking at the gene that you chose, and the research behind it, why do you think the TWI7 will work this time? Did you find something in the previous research that you would do differently?
In previous research, specifically, the research conducted by Suzanne Lee, showed that the knockout of the Twi7 gene did not lead to DNA damage accumulation. Based off of the research done on the RNAi pathway and sRNAs, it was still expected that the expression of our gene would increase after induced DNA. Using this information and our results, it makes sense that Twi7 likely does not play a role in the DNA damage response pathway in Tetrahymena, as there was no increased expression after DNA damage and when it is not present or inactive, there isn’t increased DNA damage. It is possible that the PCR we ran did not show the band because of its limited range, but I am leaning toward the conclusion that it simply just isn’t involved.
To be honest with you, I do not. It has already been discovered that the Twi proteins have a part/play a role in the RNAi pathway and assist sRNAs in their specialized functions. It is likely that the Twi7 proteins help regulate some sort of biological process that is not related to DNA, but more research needs to be done to determine its specific role.
curesymposium.org 
Can you elaborate on what it means to knockout a gene?
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To knock out a gene would basically mean to make it inactive or remove the gene itself so that no proteins can be synthesized from it. This allows for researchers to see how functions in the cell change without the presence of the gene in question. One way this is done is by using CRISPR-Cas9, but I am unsure of the method that was used when Dr.Lee found that the knockout of Twi7 leads to no DNA damage accumulation.
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You mentioned in your methods that you were creating gene specific primers, and that sounds really cool. How did you do that? If you have time to fit it in, you should what methods you used to make gene specific primers.
LikeLike
Yes, it actually is quite a fascinating process. In order to design our primers, we used the website called primer3plus, which allowed us to find a set of primers that would work best for the gene we have been given. And then, we used BLAST which allowed us to evaluate if the primers we chose would be found in other Tetrahymena genes. When the results showed us that they would not, and the primers hit the qualifications that they needed to, we were able to order them and they were delivered to the lab.
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What research could be done with this information in the scope of cancer and/or biomedical research?
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Many cancer therapies used today are targeting cancer cells to cause DNA damage and therefore kill the cell and make it nonfunctional. With our results, we know that the Twi7 gene likely does not play a role in the DNA damage response pathway, meaning it does not aid the cells in fixing the damaged DNA. It is possible that researchers could knock out the other Twi proteins that were found to play a role in the response pathway, but keep Twi7 and see how the cell responds. This could lead to a better understanding of how to target and kill cancer cells. In terms of future research that could be conducted based on the results that we got, studying the role of Twi7 in other parts of the cell would be the most beneficial.
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What research could be done with this information within the scope of cancer and/or biomedical research?
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What research coulld be done with this information within the scope of cancer and/or biomedical research?
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When looking at the gene that you chose, and the research behind it, why do you think the TWI7 will work this time? Did you find something in the previous research that you would do differently?
LikeLike
In previous research, specifically, the research conducted by Suzanne Lee, showed that the knockout of the Twi7 gene did not lead to DNA damage accumulation. Based off of the research done on the RNAi pathway and sRNAs, it was still expected that the expression of our gene would increase after induced DNA. Using this information and our results, it makes sense that Twi7 likely does not play a role in the DNA damage response pathway in Tetrahymena, as there was no increased expression after DNA damage and when it is not present or inactive, there isn’t increased DNA damage. It is possible that the PCR we ran did not show the band because of its limited range, but I am leaning toward the conclusion that it simply just isn’t involved.
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Uh thank you technology and so sorry for the 100 comments^^^
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Do you have any ideas about what other roles TW17 might play if it is not involved in DNA repair?
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To be honest with you, I do not. It has already been discovered that the Twi proteins have a part/play a role in the RNAi pathway and assist sRNAs in their specialized functions. It is likely that the Twi7 proteins help regulate some sort of biological process that is not related to DNA, but more research needs to be done to determine its specific role.
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