Enhancement of ion charging layers for the control of enzymatic DNA synthesis

Over the last decade, advancements in micro-chip technologies and biotechnologies have driven a boom in the area of gene synthesis. Currently, DNA synthesis relies heavily on the phosphoramidite chemistry, a costly and polluting approach. Additionally, synthesizing DNA strands exceeding 200 nucleotides is impractical due to the limited accuracy of the method. This limitation coupled with a difficulty to miniaturize the technology prevents further development of the DNA synthesis industry. In comparison, the recently developed enzymatic approaches, based on polymerases, hold many promises, but also, poses new challenges. One of the main challenges relates to the control over the synthesized DNA sequence, resulting directly from the management of the enzymatic activity of the polymerase. 

The polymerase activity depends on the presence of cation co-factors. Without those cations, the polymerase is inactive, and the synthesis cannot happen. By locally tweaking the cation concentration, it becomes possible to regulate the polymerase activity and the composition of the synthesized DNA strand. Then, the objective is to regulate the enzyme through its dependency on cations. The idea is to use the property of a battery-like thin film material that can take up and release cations. Under applied potential, the chosen material releases the cations which creates a localized concentration gradient where the enzyme will be active. 

In this master’s thesis, you will investigate the properties of this battery-like material. First, you will manufacture and characterize variant of the battery-like material. Second, you determine its ion insertion and extraction properties using electrochemical techniques. Finally, you will work on the patterning of the variants and investigate their new properties at the microscale for biochemical synthesis.