Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes

Liang, Zhen; Chen, Kunling; Li, Tingdong; Zhang, Yi; Wang, Yanpeng; Zhao, Qian; Liu, Jin-Xing; Zhang, Huawei; Liu, Cuimin; Ran, Yidong; Gao, Caixia

doi:10.1038/ncomms14261

Cited by 804 publications

(597 citation statements)

References 15 publications

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“…[18]. For species that require short days to trigger the reproductive phase, such as rice (Oryza sativa) and maize (Zea mays), the speed breeding technique could be used to promote rapid vegetative growth prior to reducing the photoperiod.Recent advances in genomic tools and resources [19][20][21][22][23] and the decreasing cost of sequencing have enabled plant researchers to shift their focus from model to crop plants. Despite such advances, the slow generation times of many crop plants continue to impose a high entry barrier.…”

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confidence: 99%

Speed breeding: a powerful tool to accelerate crop research and breeding

Watson

Ghosh

Williams

et al. 2017

Preprint

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The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand [1]. This slow improvement rate is attributed partly to the long generation times of crop plants. Here we present a method called 'speed breeding', . CC-BY 4.0 International license It is made available under a (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/161182 doi: bioRxiv preprint first posted online Jul. 9, 2017; Watson and Ghosh et al. (2017) 2 which greatly shortens generation time and accelerates breeding and research programs.Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum), and pea (Pisum sativum) and 4 generations for canola (Brassica napus), instead of 2-3 under normal glasshouse conditions. We demonstrate that speed breeding in fully-enclosed controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies, and transformation.The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent and potential for adaptation to larger-scale crop improvement programs. Cost-saving through LED supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing, and genomic selection, accelerating the rate of crop improvement.For most crop plants, the breeding of new, advanced cultivars, takes several years. Following crossing of selected parent lines, 4-6 generations of inbreeding are typically required to develop genetically stable lines for evaluation of agronomic traits and yield. This is particularly timeconsuming for field-grown crops that are often limited to only 1-2 generations per year. Here, we present flexible protocols for "speed breeding" that use prolonged photoperiods to accelerate the developmental rate of plants [2], thereby reducing generation time. We highlight the opportunity presented by speed breeding and detail protocols to inspire widespread adoption as a state-of-theart breeding and research tool.To evaluate speed breeding as a method to accelerate applied and basic research on cereal species, standard genotypes of spring bread wheat (T. aestivum), durum wheat (T. durum), barley (H. vulgare) and the model grass Brachypodium distachyon were grown in a controlled environment room with extended photoperiod (22 hours light/2 hours dark) ( Fig. 1; Methods:Speed breeding I; Supplementary Table 1). A light/dark period was chosen over a continuous photoperiod to support functional expression of circadian clock genes [3]. Growth was compare...

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mentioning

confidence: 99%

Speed breeding: a powerful tool to accelerate crop research and breeding

Watson

Ghosh

Williams

et al. 2017

Preprint

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“…3c) [62]. More recently, several studies have highlighted significant improvements in genome editing in plants using DNA-free CRISPR/Cas9 ribonucleoproteins [65,98]. Besides its multiplexing qualities, CRISPR has also shown great efficiency to integrate large pathways and Simultaneous downregulation of 6 genes for fatty alcohol production [48] libraries [38,88].…”

Section: Genome Engineeringmentioning

confidence: 99%

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

Ferreira

David

Nielsen

2018

Journal of Industrial Microbiology and Biotechnology

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Clustered regularly interspaced short palindromic repeats (CRISPR) is poised to become one of the key scientific discoveries of the twenty-first century. Originating from prokaryotic and archaeal immune systems to counter phage invasions, CRISPRbased applications have been tailored for manipulating a broad range of living organisms. From the different elucidated types of CRISPR mechanisms, the type II system adapted from Streptococcus pyogenes has been the most exploited as a tool for genome engineering and gene regulation. In this review, we describe the different applications of CRISPR/Cas9 technology in the industrial biotechnology field. Next, we detail the current status of the patent landscape, highlighting its exploitation through different companies, and conclude with future perspectives of this technology.

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“…One of the major limitations of using transgenic approaches to manipulate agronomically relevant traits are the associated regulatory constraints. To overcome this, the nuclease transgene can be segregated away from the edited gene(s) in subsequent generations and studies have also documented methods of Cas9-editing in wheat that avoid transgene integration altogether (Liang et al, 2017).…”

Section: Transgenic Approachesmentioning

confidence: 99%

“…For CRISPR and other transgenic approaches several varieties may be used, although only a few wheat varieties have high enough transformation efficiencies to be practical. This means that most transgenic studies in wheat are limited to a few varieties, such as Fielder, Cadenza, Bobwhite, Kenong 199 and Kronos (Li et al, 2012;Richardson et al, 2014;Liang et al, 2017). This is now changing thanks to work by groups around the world including Emma Wallington and colleagues who have expanded this portfolio to 39 varieties (http://www.niab.com/pages/id/90/crop_transformation), alongside Xingguo Ye whose group has transformed 15 Chinese varieties .…”

Section: Strategies For Usementioning

confidence: 99%

A roadmap for gene functional characterisation in wheat

Adamski

Borrill

Brinton

et al. 2018

Preprint

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Research in Arabidopsis and other model species has uncovered mechanisms regulating important biological processes in plants. With the advent of high quality functional genomic resources in wheat it is now possible to use this knowledge for crop improvement directly in wheat. In the current review, we describe some of the recent developments in wheat genomics focussing on published and publicly available resources and tools. We lay out a roadmap on how to make use of them and include a case study to exemplify them. We hope this review will be a helpful guide for plant scientists who already work on wheat or who are considering expanding their research into wheat.

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Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes

Cited by 804 publications

References 15 publications

Speed breeding: a powerful tool to accelerate crop research and breeding

Speed breeding: a powerful tool to accelerate crop research and breeding

Advancing biotechnology with CRISPR/Cas9: recent applications and patent landscape

A roadmap for gene functional characterisation in wheat

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