Authors:

Emma Scurek

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About Paper:

MYC is an oncogene that is over-expressed in the majority of cancers and helps drive cancer progression. In the promoter region of MYC there is a G-quadruplex which forms and acts as a transcriptional silencer. G- quadruplexes are noncanonical, four-stranded, secondary structures that form in guanine-rich regions of DNA. Indenoisoquinolines were originally developed as topoisomerase I inhibitors, however, our lab has previously shown that they also bind and stabilize the MYC G-quadruplex and inhibit MYC. The goal of this project is to determine the binding affinity of indenoisoquinoline compounds to the MYC promotor G-quadruplex using fluorescence spectroscopy. Fluorescence titrations are a commonly used method for determining binding affinity due to the high sensitivity of fluorescence. I used fluorophore-labeled MYC G-quadruplex DNA as a probe. I titrated indenoisoquinoline compounds into the probe. The changes in the fluorescence emissions were measured to calculate the dissociation constant (Kd) of each compound. I titrated, in duplicate, each indenoisoquinoline compound to the probe to measure binding affinity. I was able to successfully replicate both my own data as well as our lab's previous data, allowing for a more accurate comparison of the binding affinities of the different analogs. This data will be used in combination with other biophysical and cellular studies to further understanding of MYC G-quadruplex binding and stabilization and MYC downregulation. This information will help us in the development of indenoisoquinoline compounds as MYC G-quadruplex targeted anti-cancer agents.

Source:

Purdue University / 2023

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Co-authors:

Emma Scurek