GondenBraid2.0: A comprehensive toolkit for Plant Synthetic Biology

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Alejandro Sarrion-Perdigones, Marta Vazquez-Vilar M, Jorge Palaci, Antonio Granell and Diego Orzaez.

Instituto de Biologa Molecular y Celular de Plantas, Spain

Synthetic Biology has recently arrived to Plant Science, representing an emergent discipline. Following the work previously done in bacteria and yeast, Plant Synthetic Biology (PSB) aims to design and construct new biological parts or systems and redesign existing ones to carry out novel tasks. The design of plants with new characters requires the existence of a versatile and efficient modular assembly system that makes affordable the combination of complex genetic structures involving multiple transcriptional units. GoldenBraid (GB) [1] was created to overcome the existing limitations and to facilitate multigene engineering. GB is an iterative cloning system for the standardized and modular assembly of reusable genetic modules. It is based on a very efficient digestion/ligation method [2,3] that turns into a routine the assembly of combinatorial constructs. GB consists of 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 new version of GoldenBraid, named GB2.0, has recently been released, , which improves the simplicity and versatility of the previous version and provides a pre-defined grammar for transcriptional unit composition. The incorporation of new DNAparts to the collection has been standardized by using the Universal Part Domesticator Vector (pUPD). The GB2.0 includes a complete kit for PSB. Most new GBparts have been generated and tested by transient transformation of N.benthamiana leaves or by stable transformation of A.thaliana. Different vector backbones, constitutive and inducible promoters, reporters, tags, resistance genes, silencing tools (amiRNA, tasiRNA, hpRNA) and other elements of interest compose this collection that aims to serve as a reference for Plant Synthetic Biologists. 1. Sarrion-Perdigones et al.(2011).PLoSOne6: e21622. 2. Engler et al.(2008).PLoSOne3: e3647. 3. Engler et al.(2009).PLoSOne4: e5553.