Title (eng)
Generating genetically modified skin cell lines to investigate the function of LRIG3 and DIRAS1 in skin cancer
Author
Greta Schödl
Description (eng)
Bachelor thesis - University of Veterinary Medicine Vienna - 2023
Abstract (eng)
Skin cancer is the most prevalent form of cancer worldwide, with both melanoma and non-melanoma skin cancer cases increasing in recent years. GTP-binding protein Di-Ras 1 (DIRAS1) and leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) play major role in tumor development, but only little is known about functions of these proteins in skin cancer. Our previous investigations discovered that DIRAS1 and LRIG3 were upregulated in the A431 (squamous cell carcinoma) and A375 (melanoma) cell lines. From these findings, we hypothesized that DIRAS1 and LRIG3 may have tumorigenic functions in skin cancer. To gain further insights into the role of DIRAS1 and LRIG3 in skin cancer development, we planned to perform gene expression manipulation in three different skin cell lines. These included two skin cancer cell lines, A375 and A431, as well as HaCaT (immortalized human keratinocytes). To investigate the function of LRIG3 and DIRAS1 in these skin cell lines, we tried to overexpress DIRAS1 and LRIG3 genes in all three cell lines. In addition, we planned to delete DIRAS1 and LRIG3 genes by using clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 technology to generate knockout cell lines for both genes. However, we were not able to establish an overexpressing vector for DIRAS1 due to the obstacle in amplifying DIRAS1 cDNA. Moreover, in none of the cell lines, the LRIG3 gene was efficiently deleted. We also faced a problem to obtain single cell colonies of DIRAS1-knockout (KO) HaCaT cells due to their inherent difficulty in transfection. However, we successfully generated LRIG3-overexpressing A375, A431, and HaCaT cells. Likewise, we accomplished to produce DIRAS1-KO A375 and A431 cell lines. The fundamental hallmarks of cancer such as proliferation, migration as well as invasion will be analyzed in the present genetically manipulated cell lines.
Description (deu)
Bachelorarbeit - Veterinärmedizinische Universität Wien - 2023
Type (eng)
Language
[eng]
Persistent identifier
https://phaidra.vetmeduni.ac.at/o:3942
AC number
Number of pages
40
Date issued
2023
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Persistent identifier
https://phaidra.vetmeduni.ac.at/o:3942
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PDFDocument
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application/pdf
application/pdf
Created
18.03.2025 09:32:58
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