Establishment of a TP53 R248W Knock-In H460 Cell Model for the Study of CD320 Expression

Abstract

p53 is an important tumor suppressor protein that protects cells from DNA damage and cancer. When it is mutated, it loses its normal function and can even gain new, tumor-promoting functions. One such gain-of-function mutation is R248W. It can activate genes that help cancer cells grow, survive, and change their metabolism. Vitamin B12 (VB12) is a nutrient needed for DNA synthesis, metabolism, and energy production. Cells take up VB12 through the receptor CD320. Cancer cells divide quickly and need large amounts of VB12, so many tumors increase CD320 expression to bring in more VB12. Whether the R248W mutant p53 can increase CD320 expression remains an open question. To investigate this, a cellular model carrying the R248W mutation is required. In this thesis project, I aimed to establish an H460 lung cancer cell line with a TP53 R248W knock-in mutation using CRISPR-Cas9.