Grant: Undergraduate: Summer Studentship Award - Institute Community Support

In this project, the intern expands upon his in silico drug docking experience and examine a newly developed homology model. The first part of the project will allow the intern to utilize his computational skills to make mutagenesis, drug dockings and energetic analysis in a similar fashion as his previous experiment. The intern will also generate mutant SCN1A ion channels in the custom ion channel vector pICJAZZ (AES 2016 abstract # 3.076) for in vitro characterization via patch clamp electrophysiology in an in vitro expression system. Here, the intern will identify key mutations, perform primer design using Geneious software, generate mutation containing DNA via overlapping PCR followed by subsequent restriction enzyme digestion, ligation and transformation of the now mutated pICJAZZ vector. Positive colonies will be identified by Crack PCR, confirmed by Sanger Sequencing and cultured to produce sufficient mutant plasmid DNA for transient lipofectamine transfection into HEK293 cells followed by electrophysiological and pharmacological characterization using an Axopatch700B amplifier and pCLAMP10 software. Ultimately, these results will lead to improved molecular diagnostic to identify using computational models and subsequently select appropriate therapeutic agents at the time of clinical diagnosis.