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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Schaumberg, Katherine A. Antunes, Mauricio S. Kassaw, Tessema K. Xu, Wenlong Zalewski, Christopher S. Medford, June I. Prasad, Ashok |
| Description | Country affiliation: United States Author Affiliation: Schaumberg KA ( School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.); Antunes MS ( Department of Biology, Colorado State University, Fort Collins, Colorado, USA.); Kassaw TK ( Department of Biology, Colorado State University, Fort Collins, Colorado, USA.); Xu W ( Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado, USA.); Zalewski CS ( Department of Biology, Colorado State University, Fort Collins, Colorado, USA.); Medford JI ( Department of Biology, Colorado State University, Fort Collins, Colorado, USA.); Prasad A ( School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.) |
| Abstract | 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. |
| File Format | HTM / HTML |
| ISSN | 15487091 |
| Issue Number | 1 |
| Journal | Nature Methods |
| Volume Number | 13 |
| e-ISSN | 15487105 |
| Language | English |
| Publisher | Nature Publishing Group |
| Publisher Date | 2016-01-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Plants Genetics Synthetic Biology Methods Stochastic Processes Journal Article Research Support, U.s. Gov't, Non-p.h.s. |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology Biotechnology |
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