Informace o publikaci
Genome-wide CRISPR/Cas9 knockout screening revealed genes involved in CD20 regulation.
| Autoři | |
|---|---|
| Rok publikování | 2023 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Přiložené soubory | |
| Popis | CD20 antigen has been used as a target of monoclonal antibodies (mAb) such as rituximab (RTX) in the therapy of B-cell malignancies for more than two decades. However, malignant B cells downregulate CD20 on their surface, resulting in mAb resistance and therapy failure. Therefore, it is crucial to investigate the CD20 regulation to enhance the efficacy of anti CD20 mAb. This project aimed to perform CRISPR/Cas9 knockout screening to identify genes whose disruption restores CD20 surface expression. To create a model mimicking the situation in patients who have developed resistance to mAb therapy, we generated RTX-resistant CD20-low B-cell line by chronic exposure to rituximab. These cells were transduced by the GeCKO lentiviral library to obtain a collection of single-gene knockouts. After 2.5-week cultivation, the top 5% of cells with the highest expression of CD20 were sorted out. Using next-generation sequencing, we identified gene knockouts responsible for CD20 upregulation. |
| Související projekty: |