Genetic and Chemical Modifications of SP1 Nanopores

Nanopore sequencing is built upon the premise of a nanometer-sized pore penetrating either a biological or solid-state membrane, to which a voltage is applied. This electric field subsequently translates DNA strands through the nanopore, which alters the electric field according to the varying DNA base pairs passing through. There are currently a few challenges in the usage of nanopores as a viable replacement for current sequencing techniques, largely related to data resolution, accuracy, and stability. Stable Protein 1 (SP1) is a potential novel protein that addresses a few of these issues in nanopore applications due to its unique structure. SP1 is a ring-like protein complex, self-assembled from 12 identical monomers, with its oligomeric form displaying exceptional symmetry, appropriate diameter size for DNA translocation, stability, resistance to high temperatures and proteases, pH swings, detergents and organic solvents. These inherent advantages translate to SP1`s potential use as a commercial nanopore protein, which would require it to be accurate and resistant to a variety of environmental conditions and changes, among many things. Previous works done demonstrated the ability of SP1 to act as an electrode and functioning pore, as well as its ability to detect DNA molecules translocated through the protein. The aim of this project is to build upon previous works, proceeding in a fashion that aims to characterize SP1 and its derived genetic and chemical modifications in a context relevant to its use as a commercial biological nanopore protein – improving pore insertion rates, membrane integration, and ultimately sequencing accuracy.