Journal ArticleDOI
A Golden Gate Modular Cloning Toolbox for Plants
Carola Engler,Mark Youles,Ramona Gruetzner,Tim-Martin Ehnert,Stefan Werner,Jonathan D. G. Jones,Nicola J. Patron,Sylvestre Marillonnet +7 more
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TLDR
A versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable Golden Gate construction of multigene constructs for plant transformation is presented.Abstract:
Plant Synthetic Biology requires robust and efficient methods for assembling multigene constructs. Golden Gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. We present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable Golden Gate construction of multigene constructs for plant transformation. Parts include promoters, untranslated sequences, reporters, antigenic tags, localization signals, selectable markers, and terminators. The comparative performance of parts in the model plant Nicotiana benthamiana is discussed.read more
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A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
TL;DR: A versatile engineering platform for yeast, which contains both a rapid, modular assembly method and a basic set of characterized parts, and genome-editing tools for making modifications directly to the yeast chromosomes, which the authors find preferable to plasmids due to reduced variability in expression.
Journal ArticleDOI
Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease
Tom Lawrenson,Oluwaseyi Shorinola,Nicola Stacey,Chengdao Li,Lars Østergaard,Nicola J. Patron,Cristobal Uauy,Wendy Harwood +7 more
TL;DR: The use of RNA-guided Cas9 is demonstrated to generate mutations in target genes of both barley and B. oleracea and show stable transmission of these mutations thus establishing the potential for rapid characterisation of gene function in these species.
Journal ArticleDOI
NLR network mediates immunity to diverse plant pathogens.
Chih-Hang Wu,Ahmed Abd-El-Haliem,Tolga O. Bozkurt,Tolga O. Bozkurt,Khaoula Belhaj,Ryohei Terauchi,Jack H. Vossen,Sophien Kamoun +7 more
TL;DR: A complex NLR immune network is discovered in which helper NLRs in the NRC (NLR required for cell death) family are functionally redundant but display distinct specificities toward different sensor NLRs that confer immunity to oomycetes, bacteria, viruses, nematodes, and insects.
Journal ArticleDOI
Feeding the world: improving photosynthetic efficiency for sustainable crop production
TL;DR: The evidence to date that manipulation of the Calvin–Benson cycle, photorespiration, and electron transport can lead to improvement in biomass and seed yield is explored.
Journal ArticleDOI
LOCALIZER: subcellular localization prediction of both plant and effector proteins in the plant cell
Jana Sperschneider,Ann-Maree Catanzariti,Kathleen D. DeBoer,Benjamin Petre,Donald M. Gardiner,Karam B. Singh,Peter N. Dodds,Jennifer M. Taylor +7 more
TL;DR: LOCALIZER is the first method for predicting effector localisation in plants and is a valuable tool for prioritizing effector candidates for functional investigations.
References
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Journal ArticleDOI
A one pot, one step, precision cloning method with high throughput capability.
TL;DR: A cloning strategy called ‘Golden Gate’ cloning was devised that allows to obtain in one tube and one step close to one hundred percent correct recombinant plasmids after just a 5 minute restriction-ligation, thus providing precision for this fundamental process of genetic manipulation.
Journal ArticleDOI
A Modular Cloning System for Standardized Assembly of Multigene Constructs
TL;DR: A hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences.
Journal ArticleDOI
Engineering BioBrick vectors from BioBrick parts
TL;DR: The principles of part reuse and standardization are extended to BioBrick vectors so that myriad new BioBricks vectors can be readily produced from all existing and newly designed BioB Brick parts.
Journal ArticleDOI
BglBricks: A flexible standard for biological part assembly
JChristopher Anderson,John E. Dueber,Mariana Leguia,Gabriel C. Wu,Jonathan A. Goler,Jonathan A. Goler,Adam P. Arkin,Jay D. Keasling +7 more
TL;DR: The BglBrick standard provides a new, more flexible platform from which to generate standard biological parts and automate DNA assembly, and is demonstrated in three distinct applications, including the construction of constitutively active gene expression devices with a wide range of expression profiles.
Journal ArticleDOI
GreenGate---a novel, versatile, and efficient cloning system for plant transgenesis.
Athanasios Lampropoulos,Zoran Sutikovic,Christian Wenzl,Ira Maegele,Jan U. Lohmann,Joachim Forner +5 more
TL;DR: GreenGate cloning is simple and efficient since it uses only one type IIS restriction endonuclease, depends on only six types of insert modules, but at the same time allows assembling several expression cassettes in one binary destination vector from a collection of pre-cloned building blocks.