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CRISPR wheat

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https://www.readbyqxmd.com/read/29807545/expanded-base-editing-in-rice-and-wheat-using-a-cas9-adenosine-deaminase-fusion
#1
Chao Li, Yuan Zong, Yanpeng Wang, Shuai Jin, Dingbo Zhang, Qianna Song, Rui Zhang, Caixia Gao
Nucleotide base editors in plants have been limited to conversion of cytosine to thymine. Here, we describe a new plant adenine base editor based on an evolved tRNA adenosine deaminase fused to the nickase CRISPR/Cas9, enabling A•T to G•C conversion at frequencies up to 7.5% in protoplasts and 59.1% in regenerated rice and wheat plants. An endogenous gene is also successfully modified through introducing a gain-of-function point mutation to directly produce an herbicide-tolerant rice plant. With this new adenine base editing system, it is now possible to precisely edit all base pairs, thus expanding the toolset for precise editing in plants...
May 29, 2018: Genome Biology
https://www.readbyqxmd.com/read/29731763/magnesium-increases-homoeologous-crossover-frequency-during-meiosis-in-zip4-ph1-gene-mutant-wheat-wild-relative-hybrids
#2
María-Dolores Rey, Azahara C Martín, Mark Smedley, Sadiye Hayta, Wendy Harwood, Peter Shaw, Graham Moore
Wild relatives provide an important source of useful traits in wheat breeding. Wheat and wild relative hybrids have been widely used in breeding programs to introduce such traits into wheat. However, successful introgression is limited by the low frequency of homoeologous crossover (CO) between wheat and wild relative chromosomes. Hybrids between wheat carrying a 70 Mb deletion on chromosome 5B ( ph1b ) and wild relatives, have been exploited to increase the level of homoeologous CO, allowing chromosome exchange between their chromosomes...
2018: Frontiers in Plant Science
https://www.readbyqxmd.com/read/29723918/genotyping-genome-edited-mutations-in-plants-using-crispr-ribonucleoprotein-complexes
#3
Zhen Liang, Kunling Chen, Yan Yan, Yi Zhang, Caixia Gao
Despite the great achievements in genome editing, accurately detecting mutations induced by sequence-specific nucleases is still a challenge in plants, especially in polyploidy plants. An efficient detection method is particularly vital when the mutation frequency is low or when a large population needs to be screened. Here, we applied purified CRISPR ribonucleoprotein complexes to cleave PCR products for genome-edited mutation detection in hexaploid wheat and diploid rice. We show that this mutation detection method is more sensitive than Sanger sequencing and more applicable than PCR/RE method without the requirement for restriction enzyme site...
May 3, 2018: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/29710373/transforming-plant-biology-and-breeding-with-crispr-cas9-cas12-and-cas13
#4
REVIEW
Patrick Schindele, Felix Wolter, Holger Puchta
Currently, biology is revolutionized by ever growing applications of the CRISPR/Cas system. As discussed in this Review, new avenues have opened up for plant research and breeding by the use of the sequence-specific DNases Cas9 and Cas12 (formerly named Cpf1) and, more recently, the RNase Cas13 (formerly named C2c2). Although double strand break-induced gene editing based on error-prone nonhomologous end joining has been applied to obtain new traits, such as powdery mildew resistance in wheat or improved pathogen resistance and increased yield in tomato, improved technologies based on CRISPR/Cas for programmed change in plant genomes via homologous recombination have recently been developed...
April 30, 2018: FEBS Letters
https://www.readbyqxmd.com/read/29695804/targeted-mutagenesis-in-wheat-microspores-using-crispr-cas9
#5
Pankaj Bhowmik, Evan Ellison, Brittany Polley, Venkatesh Bollina, Manoj Kulkarni, Kaveh Ghanbarnia, Halim Song, Caixia Gao, Daniel F Voytas, Sateesh Kagale
CRISPR/Cas9 genome editing is a transformative technology that will facilitate the development of crops to meet future demands. However, application of gene editing is hindered by the long life cycle of many crop species and because desired genotypes generally require multiple generations to achieve. Single-celled microspores are haploid cells that can develop into double haploid plants and have been widely used as a breeding tool to generate homozygous plants within a generation. In this study, we combined the CRISPR/Cas9 system with microspore technology and developed an optimized haploid mutagenesis system to induce genetic modifications in the wheat genome...
April 25, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29642627/a-review-of-the-interactions-between-wheat-and-wheat-pathogens-zymoseptoria-tritici-fusarium-spp-and-parastagonospora-nodorum
#6
REVIEW
Adrian Duba, Klaudia Goriewa-Duba, Urszula Wachowska
Zymoseptoria tritici is a hemibiotrophic pathogen which causes Septoria leaf blotch in wheat. The pathogenesis of the disease consists of a biotrophic phase and a necrotrophic phase. The pathogen infects the host plant by suppressing its immune response in the first stage of infection. Hemibiotrophic pathogens of the genus Fusarium cause Fusarium head blight, and the necrotrophic Parastagonospora nodorum is responsible for Septoria nodorum blotch in wheat. Cell wall-degrading enzymes in plants promote infections by necrotrophic and hemibiotrophic pathogens, and trichothecenes, secondary fungal metabolites, facilitate infections caused by fungi of the genus Fusarium ...
April 10, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29388938/genome-editing-of-bread-wheat-using-biolistic-delivery-of-crispr-cas9-in-vitro-transcripts-or-ribonucleoproteins
#7
Zhen Liang, Kunling Chen, Yi Zhang, Jinxing Liu, Kangquan Yin, Jin-Long Qiu, Caixia Gao
This protocol is an extension to: Nat. Protoc. 9, 2395-2410 (2014); doi:10.1038/nprot.2014.157; published online 18 September 2014In recent years, CRISPR/Cas9 has emerged as a powerful tool for improving crop traits. Conventional plant genome editing mainly relies on plasmid-carrying cassettes delivered by Agrobacterium or particle bombardment. Here, we describe DNA-free editing of bread wheat by delivering in vitro transcripts (IVTs) or ribonucleoprotein complexes (RNPs) of CRISPR/Cas9 by particle bombardment...
March 2018: Nature Protocols
https://www.readbyqxmd.com/read/28921815/low-gluten-nontransgenic-wheat-engineered-with-crispr-cas9
#8
Susana Sánchez-León, Javier Gil-Humanes, Carmen V Ozuna, María J Giménez, Carolina Sousa, Daniel F Voytas, Francisco Barro
Coeliac disease is an autoimmune disorder triggered in genetically predisposed individuals by the ingestion of gluten proteins from wheat, barley and rye. The α-gliadin gene family of wheat contains four highly stimulatory peptides, of which the 33-mer is the main immunodominant peptide in patients with coeliac. We designed two sgRNAs to target a conserved region adjacent to the coding sequence for the 33-mer in the α-gliadin genes. Twenty-one mutant lines were generated, all showing strong reduction in α-gliadins...
April 2018: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28918562/crispr-cas9-genome-editing-in-wheat
#9
Dongjin Kim, Burcu Alptekin, Hikmet Budak
Genome editing has been a long-term challenge for molecular biology research, particularly for plants possess complex genome. The recently discovered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a versatile tool for genome editing which enables editing of multiple genes based on the guidance of small RNAs. Even though the efficiency of CRISPR/Cas9 system has been shown with several studies from diploid plants, its application remains a challenge for plants with polyploid and complex genome...
January 2018: Functional & Integrative Genomics
https://www.readbyqxmd.com/read/28913802/design-and-assembly-of-crispr-cas9-reagents-for-gene-knockout-targeted-insertion-and-replacement-in-wheat
#10
Tomáš Čermák, Shaun J Curtin
Advances in cereal transformation along with the completion of the wheat genome sequence assembly have increased the demand for tools that perform targeted and specific modifications in this crop plant. This protocol demonstrates the construction of reagents using a comprehensive genome engineering kit to create single and multiple gene "knockouts," site-specific chromosome deletions and gene replacement or "knockins" including the use of geminivirus replicons (GVRs). The reagents allow for both easy construction of simple genome engineering vectors, and "mix and match" swapping of components such as the Cas9, guide RNA and donor template cassettes for gene targeting...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28913801/targeted-mutagenesis-in-hexaploid-bread-wheat-using-the-talen-and-crispr-cas-systems
#11
Yanpeng Wang, Yuan Zong, Caixia Gao
The use of sequence-specific transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats-associated system (CRISPR/Cas9) have provided powerful reverse genetic approaches to the targeted modification of genomes in numerous organisms. Both systems have been employed to generate loss-of-function alleles in bread wheat, by targeting multiple and single copies of genes. Here we present protocols for modifying the wheat genome using the two systems. The protocols include the design of TALEN and CRISPR/Cas9 target sites and their construction, evaluation of their activities in protoplasts, transformation of plants, and mutation screening...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28913793/overview-of-the-wheat-genetic-transformation-and-breeding-status-in-china
#12
Jiapeng Han, Xiaofen Yu, Junli Chang, Guangxiao Yang, Guangyuan He
In the past two decades, Chinese scientists have achieved significant progress on three aspects of wheat genetic transformation. First, the wheat transformation platform has been established and optimized to improve the transformation efficiency, shorten the time required from starting of transformation procedure to the fertile transgenic wheat plants obtained as well as to overcome the problem of genotype-dependent for wheat genetic transformation in wide range of wheat elite varieties. Second, with the help of many emerging techniques such as CRISPR/cas9 function of over 100 wheat genes has been investigated...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28712501/gene-editing-in-polyploid-crops-wheat-camelina-canola-potato-cotton-peanut-sugar-cane-and-citrus
#13
REVIEW
Donald P Weeks
Polyploid crops make up a significant portion of the major food and fiber crops of the world and include wheat, potato, cotton, apple, peanut, citrus, and brassica oilseeds such as rape, canola, and Camelina. The presence of three sets of chromosomes in triploids, four sets in tetraploids, and six sets in hexaploids present significant challenges to conventional plant breeding and, potentially, to efficient use of rapidly emerging gene and genome-editing systems such as zinc finger nucleases, single-stranded oligonucleotides, TALE effector nucleases, and clustered regularly interspaced short palindromic repeats (CRISPR/Cas9)...
2017: Progress in Molecular Biology and Translational Science
https://www.readbyqxmd.com/read/28522548/a-multipurpose-toolkit-to-enable-advanced-genome-engineering-in-plants
#14
Tomáš Čermák, Shaun J Curtin, Javier Gil-Humanes, Radim Čegan, Thomas J Y Kono, Eva Konečná, Joseph J Belanto, Colby G Starker, Jade W Mathre, Rebecca L Greenstein, Daniel F Voytas
We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions...
June 2017: Plant Cell
https://www.readbyqxmd.com/read/28502081/simultaneous-modification-of-three-homoeologs-of-taedr1-by-genome-editing-enhances-powdery-mildew-resistance-in-wheat
#15
Yunwei Zhang, Yang Bai, Guangheng Wu, Shenghao Zou, Yongfang Chen, Caixia Gao, Dingzhong Tang
Wheat (Triticum aestivum L.) incurs significant yield losses from powdery mildew, a major fungal disease caused by Blumeria graminis f. sp. tritici (Bgt). enhanced disease resistance1 (EDR1) plays a negative role in the defense response against powdery mildew in Arabidopsis thaliana; however, the edr1 mutant does not show constitutively activated defense responses. This makes EDR1 an ideal target for approaches using new genome-editing tools to improve resistance to powdery mildew. We cloned TaEDR1 from hexaploid wheat and found high similarity among the three homoeologs of EDR1...
August 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28256553/targeted-mutagenesis-in-the-medicinal-plant-salvia-miltiorrhiza
#16
Bin Li, Guanghong Cui, Guoan Shen, Zhilai Zhan, Luqi Huang, Jiachun Chen, Xiaoquan Qi
CRISPR/Cas9 is a powerful genome editing tool that has been extensively used in model plants and crops, such as Arabidopsis thaliana, rice, wheat, and soybean. Here, we report the use of CRISPR/Cas9 to precisely knock out the committed diterpene synthase gene (SmCPS1) involved in tanshinone biosynthesis in Salvia miltiorrhiza, a traditional Chinese medicinal herb with significant pharmacological activities, such as vasorelaxation, protection against ischemia-reperfusion injury, and antiarrhythmic effects. Three homozygous and eight chimeric mutants were obtained from 26 independent transgenic hairy root lines by Agrobacterium rhizogenes-mediated transformation...
March 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28245842/knocking-out-consumer-concerns-and-regulator-s-rules-efficient-use-of-crispr-cas-ribonucleoprotein-complexes-for-genome-editing-in-cereals
#17
LETTER
Felix Wolter, Holger Puchta
Selection-free genome editing using Cas9 ribonucleoprotein embryo bombardment has been achieved for maize and wheat. This is a breakthrough that should make new breeding technologies more acceptable for worldwide use.
February 28, 2017: Genome Biology
https://www.readbyqxmd.com/read/28244994/precise-base-editing-in-rice-wheat-and-maize-with-a-cas9-cytidine-deaminase-fusion
#18
Yuan Zong, Yanpeng Wang, Chao Li, Rui Zhang, Kunling Chen, Yidong Ran, Jin-Long Qiu, Daowen Wang, Caixia Gao
Targeted base editing in plants without the need for a foreign DNA donor or double-stranded DNA cleavage would accelerate genome modification and breeding in a wide array of crops. We used a CRISPR-Cas9 nickase-cytidine deaminase fusion to achieve targeted conversion of cytosine to thymine from position 3 to 9 within the protospacer in both protoplasts and regenerated rice, wheat and maize plants at frequencies of up to 43.48%.
May 2017: Nature Biotechnology
https://www.readbyqxmd.com/read/28098143/efficient-dna-free-genome-editing-of-bread-wheat-using-crispr-cas9-ribonucleoprotein-complexes
#19
Zhen Liang, Kunling Chen, Tingdong Li, Yi Zhang, Yanpeng Wang, Qian Zhao, Jinxing Liu, Huawei Zhang, Cuimin Liu, Yidong Ran, Caixia Gao
Substantial efforts are being made to optimize the CRISPR/Cas9 system for precision crop breeding. The avoidance of transgene integration and reduction of off-target mutations are the most important targets for optimization. Here, we describe an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs). Starting from RNP preparation, the whole protocol takes only seven to nine weeks, with four to five independent mutants produced from 100 immature wheat embryos. Deep sequencing reveals that the chance of off-target mutations in wheat cells is much lower in RNP mediated genome editing than in editing with CRISPR/Cas9 DNA...
January 18, 2017: Nature Communications
https://www.readbyqxmd.com/read/27943461/high-efficiency-gene-targeting-in-hexaploid-wheat-using-dna-replicons-and-crispr-cas9
#20
Javier Gil-Humanes, Yanpeng Wang, Zhen Liang, Qiwei Shan, Carmen V Ozuna, Susana Sánchez-León, Nicholas J Baltes, Colby Starker, Francisco Barro, Caixia Gao, Daniel F Voytas
The ability to edit plant genomes through gene targeting (GT) requires efficient methods to deliver both sequence-specific nucleases (SSNs) and repair templates to plant cells. This is typically achieved using Agrobacterium T-DNA, biolistics or by stably integrating nuclease-encoding cassettes and repair templates into the plant genome. In dicotyledonous plants, such as Nicotinana tabacum (tobacco) and Solanum lycopersicum (tomato), greater than 10-fold enhancements in GT frequencies have been achieved using DNA virus-based replicons...
March 2017: Plant Journal: for Cell and Molecular Biology
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