Proteomic Puzzles: Investigating the Post-Translational Modifications of Sox2 — 67a — Calla Freesemann, Ashley Nettifee             

Small Cell Lung Cancer (SCLC) is a rare form of cancer with notable characteristics of low survival rates, accelerated metastasis, and chemoresistance after an initially successful trial of chemotherapy. One noteworthy aspect of SCLC that previous studies have identified is the protein Sox2, which was discovered to be a driver of SCLC, one that aids in the initiation and growth of tumors. Proteins such as Sox2 are specialized and diversified when they undergo post-translational modifications (PTMs). By identifying the PTMs of Sox2, scientists will be able to take note of the kinds of alterations being made to the cells in SCLC, further improving their knowledge of the cancer’s inner workings. Despite its deadly nature, remarkably few sustainable therapies exist to treat SCLC. In order to better the chances of understanding SCLC, it is necessary to further investigate its underlying mechanisms. By researching the PTMs of Sox2, scientists will be able to devise specialized and sustainable treatments for better patient outcomes. In order to identify these modifications, Sox2 must be tagged and isolated, which will come to fruition through the use of techniques such as nucleofection, sorting, selection, pull-down, and mass spectrometry. A FLAG tag will be inserted into the genome of SCLC cells, at the C-terminus of Sox2, then the cells will be sorted into single cells that will be grown up and then selected using DNA extractions and PCR. Cell samples that successfully inserted the FLAG tag will be identified, pulled down, and sent off for mass spectrometry, which would ultimately classify the post-translational modifications of Sox2. Currently, validation is being done to assess whether or not a successful FLAG tag insert is present in these clones in order to further the breadth of knowledge about Sox2 and ultimately increase the regrettably low survival rate of SCLC patients.

Sanford Research
Dr. Michael Kareta