DNA damage induces cell cycle arrest, required for the action of repair mechanisms. The genes of these pathways affect cells' sensitivity to DNA damage, and their loss may lead to deregulated cellular growth, genomic instability and cancer. Until now, our knowledge in this field was gained mainly from work in cells of DNA damage-sensitive individuals and from lower eukaryotes. Improving our understanding of these processes in mammals was hampered due to lack of suitable genetic tools. Recently, we developed a vector-based system that generates persistent loss-of-function phenotypes in mammalian cells by production of short interfering RNA (siRNA). A siRNA-expressing library to suppress up to 8000 human genes was constructed in our institute and is consent to perform genome wide loss-of-function screens.
The proposal includes:
1) Screen for novel genes participating in DNA damage response pathways;
2) Elucidating the roles of these genes in DNA damage response pathways;
3) Evaluating the contribution of these genes to malignancy.
I will screen primary human cells for genes involved in sensitising cells to genotoxic stress, and genes that synergize with radio- and chemotherapy agents in tumour cell killing. I will investigate the mechanism of action of the identified genes in DNA damage response pathways, and will assess their tumour suppressive/oncogenic properties. The identification of novel genes modulating cellular toxicity induced by DNA damage will improve our understanding of the action genotoxic agents, and will provide new opportunities to screen for drugs that sensitise tumours to conventional therapy.
This project will render the siRNA library system extremely beneficial for mammalian models and research fields, encouraging the transfer of research competencies across different labs. My training in this project will complement my contemporary expertise and will diversify them considerably, contributing to my development as an independent researcher.
Fields of science
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