GB3.0: a platform for plant bio-design that connects functional DNA elements with associated biological data

Nucleic Acids Res. 2017 Feb 28;45(4):2196-2209. doi: 10.1093/nar/gkw1326.

Abstract

Modular DNA assembly simplifies multigene engineering in Plant Synthetic Biology. Furthermore, the recent adoption of a common syntax to facilitate the exchange of plant DNA parts (phytobricks) is a promising strategy to speed up genetic engineering. Following this lead, here, we present a platform for plant biodesign that incorporates functional descriptions of phytobricks obtained under pre-defined experimental conditions, and systematically registers the resulting information as metadata for documentation. To facilitate the handling of functional descriptions, we developed a new version (v3.0) of the GoldenBraid (GB) webtool that integrates the experimental data and displays it in the form of datasheets. We report the use of the Luciferase/Renilla (Luc/Ren) transient agroinfiltration assay in Nicotiana benthamiana as a standard to estimate relative transcriptional activities conferred by regulatory phytobricks, and show the consistency and reproducibility of this method in the characterization of a synthetic phytobrick based on the CaMV35S promoter. Furthermore, we illustrate the potential for combinatorial optimization and incremental innovation of the GB3.0 platform in two separate examples, (i) the development of a collection of orthogonal transcriptional regulators based on phiC31 integrase and (ii) the design of a small genetic circuit that connects a glucocorticoid switch to a MYB/bHLH transcriptional activation module.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology / methods*
  • DNA, Plant*
  • Gene Expression
  • Gene Expression Regulation, Plant
  • Genes, Reporter
  • Plants / genetics*
  • Plants / metabolism*
  • Promoter Regions, Genetic
  • Protoplasts / metabolism
  • Software*
  • Transcription, Genetic
  • User-Computer Interface
  • Web Browser

Substances

  • DNA, Plant