Question

2. You are studying a (hypothetical) newly-described member of a family of protein kinases (XYK6), which you suspect may function as an oncogene, based upon the observation that specific point mutations (especially Thr489Asp) are sometimes seen in human breast tumors. You’ve queried the TCGA database and found that this mutation, when present, seems to be an “early” mutation, since it is seen as frequently in early-stage lesions as in later-stage tumors. You therefore hypothesize that it may be a driver mutation that contributes to the process of breast cancer initiation. Describe a mouse model that would allow you to test this hypothesis.

• For xenograft/allograft experiments, briefly describe the type of cell lines you might use and the experimental end points you would be looking at.
• For GEMs, briefly describe the type of mouse you would make (you don’t need to go into details about how the mouse is generated), and the experimental end points you would be looking at.

*Please type answer, written answers are often hard to read!

Answer 1

For xenograft/allograft experiments- In this case, we can use TCP005 breast cancer cell line. There can be two approaches in which we can do the experiment- 1) we can do mutation in the cell line by CRISPER-CAS9 technique and introduce the mutation after which we will inject the mutant cell and measure the tumor size with respect to control. 2) We can delete the kinase gene and introduce the mutant kinase gene in the cell. Then we can repeat the above-said experiment.

For GEMs- We can use the nude mouse (which lacks thymus) which will lack the adaptive immune so that we can observe the effect of the mutant in the progression of the tumor. This will be independent of the host immune response. In this case also we will measure the size of the tumor in the mutant with respect to the control.