Terveer, Melanie (2022) CRISPR/Cas9 KOs of p53 and STAT1 in HEK293 Cells. Bachelor's Research Project, Life Science and Technology.
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Abstract
Chromosomal instability (CIN) is a process characterized by chromosome mis-segregation often occurring in cancer. As a result of CIN, a downstream inflammatory response is activated, which leads to cancer cell clearance. STAT1 and p53 are genes which are known to be involved in this inflammatory response, while OAS1 and IFIT1 are theorized to be involved. To better understand the role of these genes, the plan was to make knockouts using the CRISPR/Cas9 mechanism. HEK293 cells were transfected with plasmids containing sgRNAs for p53 and STAT1. Validation of the knockouts was performed, using PCR and subsequent sequencing and western blot. The western blot showed that there was no decrease in STAT1 protein levels in the STAT1 KO sample. Conversely, there was a slight reduction of p53 protein levels in the p53 KO sample, but PCR and Sanger sequencing showed that there was no mutation at the p53 sgRNA target site of p53 KO cells. Therefore, it seems that the knockout of p53 and STAT1 in HEK293 cells was unsuccessful.
| Item Type: | Thesis (Bachelor's Research Project) |
|---|---|
| Supervisor name: | Broekhuis, M.J.C. |
| Degree programme: | Life Science and Technology |
| Thesis type: | Bachelor's Research Project |
| Language: | English |
| Date Deposited: | 09 Jun 2022 09:47 |
| Last Modified: | 09 Jun 2022 09:47 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/27220 |
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