Precise control of gene expression can be used to reprogram cells from one state to another. Specifically, the reprogramming of fibroblasts to motor neurons can be used to treat injuries in the striatum and spinal cord. Achieving this level of control is difficult and requires synthetic control systems that are robust and well-tuned. The goal of my project is to validate ligand-responsive splicing switches, currently an underexplored method for controlling gene expression in reprogramming. I aim to extend the splicing switch to other fluorescent proteins and epitope tags, and to titrate and control expression levels of multiple transcription factors and genes from a single switch. Currently, we have a guanine-responsive splicing switch that is incorporated in a mCherry fluorescent protein sequence, and preliminary results show the splicing switch successfully titrates mCherry levels along a range of guanine concentrations and produces unimodal shifts in expression in HEK293Ts. Successful implementation of the splicing switch in other modalities will greatly expand the toolbox of genetic parts for the Galloway lab and enable rapid portability to other projects.

Through this SuperUROP, I hope to gain more experience in synthetic biology research as well as generally, more confidence in my scientific communication and presentation skills. I am excited to apply knowledge from various classes I have taken to continue working on this project that has possible therapeutic applications which aligns with my interests in medicine.