The Effect of STN1 Knockout on the Atr DNA Damage Repair Pathway
School Name
Ridge View High School
Grade Level
12th Grade
Presentation Topic
Cell and Molecular Biology
Presentation Type
Mentored
Abstract
This study was undertaken in order to determine the effect of the removal of STN1 on the ATR DNA damage repair pathway. STN1 is one of the structural proteins in the newly discovered protein complex CST, which consists of CTC1, STN1, and TEN1 in humans and has been found to play a role in rare genetic diseases Coats Plus and dyskeratosis congenita. As CTC1 was discovered to play a role in DNA damage repair, it was hypothesized that STN1, a related protein in the complex, may also play a related role in this pathway. In order to test this, an induced knockout of STN1 was created in a HeLa cell line using a Cas9 complex. The knockout and a wild type of this cell line were collected and frozen on even numbered days 2-12. The protein from these cells was extracted and quantified to run in a Western blotting procedure, testing for the presence of ATR downstream products and preliminary factors. The resulting blots showed an identifiable decrease in the presence of inducers early on (days 4-8), with a rescue of the tested preliminary factor in days 10 and 12. Downstream product levels were unable to be identified, as the tested cell line showed a p53 and consequently a p21 deficiency. It was thus revealed that STN1 has an effect on the ATR DNA damage repair pathway, though further research is needed to fully confirm and investigate the specific role of the STN1 protein in this pathway. Given the importance of DNA damage repair in cellular maintenance and reproduction, the understanding of specific mechanisms in this pathway opens up the possibility of specific treatments for the failures of the pathway.
Recommended Citation
Van Alstine, Ericka, "The Effect of STN1 Knockout on the Atr DNA Damage Repair Pathway" (2020). South Carolina Junior Academy of Science. 274.
https://scholarexchange.furman.edu/scjas/2020/all/274
Location
Furman Hall 107
Start Date
3-28-2020 12:15 PM
Presentation Format
Oral Only
Group Project
No
The Effect of STN1 Knockout on the Atr DNA Damage Repair Pathway
Furman Hall 107
This study was undertaken in order to determine the effect of the removal of STN1 on the ATR DNA damage repair pathway. STN1 is one of the structural proteins in the newly discovered protein complex CST, which consists of CTC1, STN1, and TEN1 in humans and has been found to play a role in rare genetic diseases Coats Plus and dyskeratosis congenita. As CTC1 was discovered to play a role in DNA damage repair, it was hypothesized that STN1, a related protein in the complex, may also play a related role in this pathway. In order to test this, an induced knockout of STN1 was created in a HeLa cell line using a Cas9 complex. The knockout and a wild type of this cell line were collected and frozen on even numbered days 2-12. The protein from these cells was extracted and quantified to run in a Western blotting procedure, testing for the presence of ATR downstream products and preliminary factors. The resulting blots showed an identifiable decrease in the presence of inducers early on (days 4-8), with a rescue of the tested preliminary factor in days 10 and 12. Downstream product levels were unable to be identified, as the tested cell line showed a p53 and consequently a p21 deficiency. It was thus revealed that STN1 has an effect on the ATR DNA damage repair pathway, though further research is needed to fully confirm and investigate the specific role of the STN1 protein in this pathway. Given the importance of DNA damage repair in cellular maintenance and reproduction, the understanding of specific mechanisms in this pathway opens up the possibility of specific treatments for the failures of the pathway.
