Kytopen: Expanding the Language of Biology with Pulsed Electric Fields

The potential to solve many of mankind's most pressing challenges including the needs for alternative fuels, enhancing oil recovery, and even treating cancer could involve engineering the numerous bacteria currently beyond one's reach. There are many types of bacteria that can be grown in the lab but their potential impact on people's lives has not been realized. New tools are needed to unlock bacteria?s true potential to solve many challenges of interest to mankind. Creating tools to accelerate discovery of new cancer treatments, alternative fuels, or low cost biomaterials will significantly impact human life. Population growth and increasing lifespans are putting ever increasing demands on the environment, driving need for alternative sources of fuels, food, and medicines. Bacteria have been a bit neglected as sources for innovation; however, they already have the internal machinery to help do industrial processes in a fast and efficient manner.

Pulsed electric fields (i.e. electroporation) can be used to deliver genetic material into many types of cells including bacteria. The goal of this proposal is to substantially expand the successful application of pulsed electric fields to bacteria and enable discovery of new applications in genetic engineering and synthetic biology. A major limitation of synthetic biology is the inability to incorporate genetic material into many bacteria due to the challenge of permeating the cell envelope while maintaining high cell viability. This team's goal is to reduce the time required to develop tools to genetically manipulate bacteria from months or even years to days. This I-Corps team has developed a proof-of-concept microfluidic device to enable the characterization of the critical electric field for bacterial electroporation under specific experimental conditions in a single experiment. The team is devising a technique to test several thousand unique conditions within a single day, spanning the entire parametric space. This will be done using a microfluidic platform, or several in parallel, to test small aliquots of cells at varying experimental conditions. Therefore, the proposed microfluidic system enables quantification of the critical electroporation parameters in a single experiment, which would otherwise require months of experimentation. The proposed innovation will allow two significant advancements: 1) the ability to quickly determine high yield electroporation conditions and 2) the ability to utilize many of the more than 10,000 bacterial strains that have been heretofore intractable or difficult to genetically manipulate.