Planta Med 2013; 79 - WS18
DOI: 10.1055/s-0033-1351820

GondenBraid2.0: A comprehensive toolkit for plant synthetic biology

A Sarrion-Perdigones 1, M Vazquez-Vilar 1, J Palaci 1, A Granell 1, D Orzáez 1
  • 1Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas. Universidad Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain

Plant Synthetic Biology (PSB) aims to design plants with novel traits by introducing new biological parts or systems or by redesigning existing ones to carry out novel tasks. This will be fostered by the employ of modular DNA assembly systems that facilitate the writing of complex genetic instructions while enabling the exchange and reuse of new synthetic modules. GoldenBraid (GB) [1] is a modular DNA assembly method created to overcome the existing limitations for multigene engineering in plants. GB iterative cloning loop is based on a very efficient digestion/ligation method [2,3] that turns into a routine the assembly of combinatorial constructs. The GB cloning kit comprises a set of eight destination plasmids (pDGBs) designed to host scar-benign multipartite composites that can be binarily combined to create complex multigene constructs. GB makes possible the assembly of 15 – 19 kb constructs comprising 4 – 5 transcription units made of individual standardized GBparts in a few days work.

A recently released new version of GoldenBraid, named GB2.0, [4] proposes a modular cloning schema with positional notation that resembles the grammar of natural languages. Use of the GB2.0 framework is facilitated by a number of web resources (www.gbcloning.org), which include a publicly available database, tutorials and a software package that provides in silico simulations and lab protocols. A growing number of vector backbones, constitutive and inducible promoters, reporters, tags, marker genes and silencing tools (amiRNA, tasiRNA, hpRNA) among other elements complete the GB2.0 framework, which aims to serve as a reference for Plant Synthetic Biologists.

References:

[1] Sarrion-Perdigones (2011). PLoSOne6: e21622.

[2] Engler (2008). PLoSOne3: e3647.

[3] Engler (2009). PLoSOne4: e5553.

[4] Sarrión-Perdigones (2012). Plant Physiol. In press.