Identification of Genetic Components Effective in Cisplatin Resistance in Non-Small Cell Lung Cancer Calu1 Cell Line by Whole Genome CRISPR/Cas9-Based Genetic Screening Method
Project no. 22673 · Status: Kapanmış (Yayın bekleniyor) · 2021
Summary
<p>Lung cancer is a common type of cancer and one of the leading causes of cancer-related death. Cisplatin is a chemotherapeutic agent widely used in the treatment of many types of cancer, including lung cancer. It is known that ~60% of cases develop resistance over time to cisplatin, which is used in the standard treatment of non-small cell lung cancer (NSCLC); In our project proposal, we aim to illuminate the mechanisms underlying this resistance. For this purpose, we aim to identify the components on which the cisplatin-sensitive and resistant NSCLC cell line Calu1 depends, using a CRISPR/Cas9-based genetic screening method, and to investigate the usability of these components in cancer treatment.</p><p>In the study, the lentiviral 'Brunello library', which targets 19114 human genes and consists of a total of 76441 guide RNAs (gRNAs), will be used for CRISPR/Cas9-based genetic screening. After Calu1 cells are made resistant to cisplatin, cisplatin-sensitive and resistant Calu 1 cells will be transfected with the library, and after 14-21 days, the gRNA distribution between the groups will be determined by bioinformatic analyzes that will be performed after next-generation sequence analysis. Here, in particular, the genes targeted by gRNAs that are decreased or increased in resistant Calu1 cells will be listed, and after the candidate genes selected among them are silenced (knock-out) in resistant Calu1 cells, their effects on important cellular functions such as cell viability, apoptosis and cell cycle will be investigated with advanced functional tests.</p><p>Our project is unique in that the genes that are effective in cisplatin resistance in the NSCLC disease model will be identified for the first time by a CRISPR/Cas9-based genetic screening method. The outputs will be used in new projects to restore the sensitivity of cancer cells to cisplatin and to identify new drug targets.</p>