Quantitative characterization of genetic parts and circuits for plant synthetic biology
| Autor: | June I. Medford, Mauricio S. Antunes, Tessema K. Kassaw, Ashok Prasad, Wenlong Xu, Katherine A Schaumberg, Christopher S Zalewski |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Normalization (statistics) Stochastic Processes biology Gene Circuits business.industry Gene regulatory network food and beverages Cell Biology Plants biology.organism_classification Biochemistry Characterization (materials science) Biotechnology 03 medical and health sciences Synthetic biology Transformation (genetics) 030104 developmental biology Arabidopsis Synthetic Biology business Biological system Molecular Biology Electronic circuit |
| Zdroj: | Nature Methods. 13:94-100 |
| ISSN: | 1548-7105 1548-7091 |
| Popis: | Plant synthetic biology promises immense technological benefits, including the potential development of a sustainable bio-based economy through the predictive design of synthetic gene circuits. Such circuits are built from quantitatively characterized genetic parts; however, this characterization is a significant obstacle in work with plants because of the time required for stable transformation. We describe a method for rapid quantitative characterization of genetic plant parts using transient expression in protoplasts and dual luciferase outputs. We observed experimental variability in transient-expression assays and developed a mathematical model to describe, as well as statistical normalization methods to account for, this variability, which allowed us to extract quantitative parameters. We characterized >120 synthetic parts in Arabidopsis and validated our method by comparing transient expression with expression in stably transformed plants. We also tested >100 synthetic parts in sorghum (Sorghum bicolor) protoplasts, and the results showed that our method works in diverse plant groups. Our approach enables the construction of tunable gene circuits in complex eukaryotic organisms. |
| Databáze: | OpenAIRE |
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