CRISPR/Cas9‐Based Multiplex Genome Editing in Monocot and Dicot Plants

Ma, Xingliang; Liu, Yao-Guang

doi:10.1002/cpmb.10

Cited by 113 publications

(84 citation statements)

References 34 publications

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“…To confirm that TCD10 was blame for the mutant phenotype observed, both RNAi and CRISPR/Cas9 genome editing technology, which is of significance for basic plant research and crop genetic improvement (Belhaj et al 2015; Ma and Liu 2016; Ma et al 2016), were preformed. In RNAi experiments, the construct vector pTCK303 with a maize ubiquitin promoter and a rice intron was used as an RNAi vector (Wang et al 2004).…”

Section: Methodsmentioning

confidence: 99%

The Rice Pentatricopeptide Repeat Gene TCD10 is Needed for Chloroplast Development under Cold Stress

Jun

Liu

et al. 2016

Rice

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BackgroundChloroplast plays a vital role in plant development and growth. The pentatricopeptide repeat (PPR) gene family is one of the largest gene families in plants. In addition, cold stress affects a broad spectrum of cellular components, e.g. chloroplast, and metabolism in plants. However, the regulatory mechanism for rice PPR genes on chloroplast development still remains elusive under cold stress.ResultIn this paper, we characterized a new rice PPR gene mutant tcd10 (thermo-sensitive chlorophyll-deficient mutant 10) that exhibits the albino phenotype, malformed chloroplast and could not survive after the 5-leaf stage when grown at 20 °C, but does the normal phenotype at 32 °C. Map-based cloning, followed by RNA interference and CRISPR/Cas9 genome editing techniques, revealed that TCD10 encoding a novel PPR protein, mainly localized to the chloroplasts, with 27 PPR motifs, is responsible for the mutant phenotype. In addition, TCD10 is specific expression in tissues. The disruption of TCD10 resulted in an evidently reduced expression of chloroplast-associated genes under cold stress (20 °C), whereas they did recovered to normal levels at high temperature (32 °C). These results showed an important role of TCD10 for chloroplast development under cold stress.ConclusionsThe TCD10 encodes a novel rice PPR protein, mainly located in chloroplasts, which is important for chloroplast development, growth and the maintenance of photosynthetic electron transport and its disorder would lead to an aberrant chloroplast and abnormal expressions in these genes for chloroplast development and photosynthesis in rice under cold stress.Electronic supplementary materialThe online version of this article (doi:10.1186/s12284-016-0134-1) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

The Rice Pentatricopeptide Repeat Gene TCD10 is Needed for Chloroplast Development under Cold Stress

Jun

Liu

et al. 2016

Rice

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“…The utility of CRISPR/Cas for targeted gene editing and GE has been proved in many studies in plants (Wang et al ., ; Bortesi and Fischer, ; Schaeffer and Nakata, ; Hilscher et al ., ; Ma et al ., ; Puchta, , ; Quetier, ; Samanta et al ., ; Schiml and Puchta, ; Steinert et al ., ). Furthermore, multiplex GE has been demonstrated in rice (Ma and Liu, ) and Arabidopsis (Xing et al ., ).…”

Section: New Tools For Precise Natural Genome Engineeringmentioning

confidence: 99%

From classical mutagenesis to nuclease‐based breeding – directing natural DNA repair for a natural end‐product

2017

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Production of mutants of crop plants by the use of chemical or physical genotoxins has a long tradition. These factors induce the natural DNA repair machinery to repair damage in an error-prone way. In the case of radiation, multiple double-strand breaks (DSBs) are induced randomly in the genome, leading in very rare cases to a desirable phenotype. In recent years the use of synthetic, site-directed nucleases (SDNs) - also referred to as sequence-specific nucleases - like the CRISPR/Cas system has enabled scientists to use exactly the same naturally occurring DNA repair mechanisms for the controlled induction of genomic changes at pre-defined sites in plant genomes. As these changes are not necessarily associated with the permanent integration of foreign DNA, the obtained organisms per se cannot be regarded as genetically modified as there is no way to distinguish them from natural variants. This applies to changes induced by DSBs as well as single-strand breaks, and involves repair by non-homologous end-joining and homologous recombination. The recent development of SDN-based 'DNA-free' approaches makes mutagenesis strategies in classical breeding indistinguishable from SDN-derived targeted genome modifications, even in regard to current regulatory rules. With the advent of new SDN technologies, much faster and more precise genome editing becomes available at reasonable cost, and potentially without requiring time-consuming deregulation of newly created phenotypes. This review will focus on classical mutagenesis breeding and the application of newly developed SDNs in order to emphasize similarities in the context of the regulatory situation for genetically modified crop plants.

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“…To knock‐out PHS9 and OsGAP , the U3 and U6a primers for PHS9 and OsGAP were mixed (Table S4) to 1 μ m , heated at 90°C for 30 s, and annealed to 25°C, and then inserted into U3 or U6a sgRNA intermediate plasmid at the Bsa I site. The gRNA expression cassettes were obtained by two rounds of PCR using the linkage products as a template, U‐F/gRNA‐R primers (first), and B1′/B2 and B2′/BL primers (second); the products were inserted into pYLCRISPR/Cas9‐MH binary vectors at Bsa I sites to obtain the complete PHS9KO and GAPKO knock‐out vectors, respectively (Ma and Liu, ).…”

Section: Methodsmentioning

confidence: 99%

Control of rice pre‐harvest sprouting by glutaredoxin‐mediated abscisic acid signaling

Tang

Gao

et al. 2019

The Plant Journal

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Pre-harvest sprouting (PHS) is one of the major problems in cereal production worldwide, which causes significant losses of both yield and quality; however, the molecular mechanism underlying PHS remains largely unknown. Here, we identified a dominant PHS mutant phs9-D. The corresponding gene PHS9 encodes a higher plant unique CC-type glutaredoxin and is specifically expressed in the embryo at the late embryogenesis stage, implying that PHS9 plays some roles in the late stage of seed development. Yeast two-hybrid screening showed that PHS9 could interact with OsGAP, which is an interaction partner of the abscicic acid (ABA) receptor OsRCAR1. PHS9or OsGAP overexpression plants showed reduced ABA sensitivity in seed germination, whereas PHS9 or OsGAP knock-out mutant plants showed increased ABA sensitivity in seed germination, suggesting that PHS9 and OsGAP acted as negative regulators in ABA signaling during seed germination. Interestingly, the germination of PHS9 and OsGAP overexpression or knock-out plant seeds was weakly promoted by H 2 O 2 , implying that PHS9 and OsGAP could affect reactive oxygen species (ROS) signaling during seed germination. These results indicate that PHS9 plays an important role in the regulation of rice PHS through the integration of ROS signaling and ABA signaling.

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CRISPR/Cas9‐Based Multiplex Genome Editing in Monocot and Dicot Plants

Cited by 113 publications

References 34 publications

The Rice Pentatricopeptide Repeat Gene TCD10 is Needed for Chloroplast Development under Cold Stress

The Rice Pentatricopeptide Repeat Gene TCD10 is Needed for Chloroplast Development under Cold Stress

From classical mutagenesis to nuclease‐based breeding – directing natural DNA repair for a natural end‐product

Control of rice pre‐harvest sprouting by glutaredoxin‐mediated abscisic acid signaling

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