The Cellular Engineering Group works to provide measurement assurance for more reproducible and comparable cell-free expression systems.
NIST has several, on-going projects in its growing portfolio to support fundamental understanding and practical applications of cell-free systems. First, the Cellular Engineering Group complements existing efforts to determine sources of variability by providing guidelines for DNA template preparation for more reproducible and comparable cell-free protein production. This study leverages automation to explore potential sources of uncertainty, as well as reduce human error. Second, integrating engineered biological parts to achieve more sophisticated functions will require investment in measurement platforms and methods. Third, the Group sees potential for integrating synthetic biology and nanotechnology to advance engineering biology. Cell-free systems lack potentially functional subcellular confinement and localization, and selective reintroduction of that confinement using nanofluidics may offer new engineering capabilities. A chip-based format also facilitates single molecule measurements of dynamic cell-free reactions to inform modeling towards predictive function and design. Fourth, developing a workflow and measurements for end-to-end automation of cell-free reactions will help the Group understand how to better serve the cell-free community and emerging related industries, for example through rapid optimization of cell-free systems derived from new organisms and a generalizable method for comprehensive characterization of cell-free reactions.
Romantseva E.F., Tack D.S., Alperovich N., Ross D., Strychalski E.A. (2022) Best Practices for DNA Template Preparation Toward Improved Reproducibility in Cell-Free Protein Production. In: Karim A.S., Jewett M.C. (eds) Cell-Free Gene Expression. Methods in Molecular Biology, vol 2433. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1998-8_1
“CELL-FREE (Comparable Living Lysates For Research, Education, and Entrepreneurship) workshop Report” E. Romantseva, E. A. Strychalski. https://doi.org/10.6028/NIST.SP.1500-13