Quantifying Genotypic Diversity Caused by Sporulation
Abstract
Engineered spore devices can be used to deploy synthetic biology circuits to remote locations without the need of cold chain storage. Army relevant applications of these could be field production of food, fuels and structural materials; battlefield production of biologic therapies; or dormant environmental sensors capable of Ôwaking upÕ and transducing information from surfaces, water supplies, etc. One limitation to these devices is the heterogeneity of their response after germinating from spore state. Although most studies attribute this purely to phenotypic causes, we have recent work that indicates there is likely a genotypic cause as well: mutation caused by low-fidelity polymerases up-regulated in late stationary growth phases (when the cell is under stress and goes into spore form). It is the purpose of this short term grant to create a reliable and fast method for screening the heterogeneity of expression cassettes in cell populations before and after sporulation. We propose to develop such a technique using nanopore chips which would enable direct reading of full length cassettes without amplification or fragmentation of the starting material. Moreover, by using this technique we will be able to compare the effect of different cell types, method of plasmid replication, and size of expression cassette. These finding and methods will enable us to embark on larger studies with Army Lab Researchers on spore based devices that are stable, robust, and house genetic circuits relevant to the Army.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jun 25, 2021
- Source ID
- W911NF2110058
Entities
People
- Nigel Reuel
Organizations
- Army Contracting Command
- Iowa State University
- United States Army