Enhancing Ministring DNA Purification

Overview

Gene therapy is a powerful technique used to treat or prevent diseases. Vectors are used in gene therapy to deliver therapeutic genes into the targeted cells. There are two types of vectors: viral and non-viral. LCC (linear covalently closed) ministring DNA belongs to the non-vector category and is synthesized in the lab using an E. coli-based recombinant platform. This method is cost-effective.

Since the synthesis produces unwanted byproducts, these byproducts need to be removed to enhance msDNA purification and require a cost-effective purification method to produce msDNA on a larger scale. This project focuses on the overexpression of the Pi-Sce1 homing endonuclease enzyme, which targets the msDNA-synthesizing precursor plasmid, digests the unwanted byproducts of msDNA synthesis. Another approach of this project is the use of the CRISPR-Cas3 system present in E. coli K-12 bacteria to digest the unwanted byproducts of msDNA synthesis.