Investigating the Functional Significance of the Novel lncRNA in Breast Cancer

Breast cancer is the most common cancer diagnosed and the leading cause of cancer-related deaths in women, posing a global health concern. Several studies have explored the influence of non-coding RNAs (ncRNAs) on the initiation and progression of cancer. Non-coding RNA (ncRNA) refers to a category of RNA molecules that do not possess the ability to encode proteins. Previous studies have indicated that long non-coding RNAs (lncRNA), which ncRNAs that are more than 200 bp long, play important roles in cancer development & progression. In addition, ncRNAs act as microRNAs (miRNAs) decoy which are small ncRNAs by competing with other molecules for binding to and sequestering miRNA. RNA I is a previously unstudied lncRNA that was recently identified as an unannotated region in the genome of MCF7 breast cancer cells, but was absent in the non-cancerous breast MCF10A cells. Given its novel status and potentially specific expression in cancer cells, this research aims to uncover the functions of RNAI, which is 5268 bp-long, and its potential interaction with miRNAs in breast cancer. This project uses a CRISPR–Cas9 genome editing approach to generate a knockout model of RNAI in MCF7 breast cancer cells. Complementarily, small interfering RNA (siRNA) will be used to transiently silence RNA I with the goal of investigating the impact of RNAI expression on breast cancer cell behavior. On establishing and validating the cell line model, assays will be conducted to assess cell proliferation (CellTitre Glo), apoptosis (Annexin V/Propidium iodide flow cytometry) and the inflammatory response (MyD88 and PD-L1 expression) in response to RNAI knockout/knockdown. In summary, this study is aimed to understand the role of RNA I in breast cancer and its potential use as an anti-cancer target, with the goal of developing novel treatment approaches for breast cancer management.