A Simple CRISPR/Cas9 System for Multiplex Genome Editing in Rice

Chün, Wang; Shen, Lan; Fu, Yaping; Yan, Changjie

doi:10.1016/j.jgg.2015.09.011

Cited by 121 publications

(78 citation statements)

References 11 publications

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“…Multiplexing greatly expands genome editing capabilities as it allows more efficient generation of large chromosomal deletion mutations and facilitates CRISPR based epigenetic genome modification (Paul and Qi, 2016; Puchta, 2016). Moreover, robust and easy to use molecular “toolkits” are available to streamline the assembly and expression of multiplexed gRNAs and support many different Cas9 variants and downstream applications (Xing et al, 2014; Lowder et al, 2015; Ma et al, 2015; Wang, C. et al, 2015; Zhang, Z. et al, 2016). …”

Section: Introductionmentioning

confidence: 99%

Rapid Evolution of Manifold CRISPR Systems for Plant Genome Editing

Lowder

Malzahn

2016

Front. Plant Sci.

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Advanced CRISPR-Cas9 based technologies first validated in mammalian cell systems are quickly being adapted for use in plants. These new technologies increase CRISPR-Cas9's utility and effectiveness by diversifying cellular capabilities through expression construct system evolution and enzyme orthogonality, as well as enhanced efficiency through delivery and expression mechanisms. Here, we review the current state of advanced CRISPR-Cas9 and Cpf1 capabilities in plants and cover the rapid evolution of these tools from first generation inducers of double strand breaks for basic genetic manipulations to second and third generation multiplexed systems with myriad functionalities, capabilities, and specialized applications. We offer perspective on how to utilize these tools for currently untested research endeavors and analyze strengths and weaknesses of novel CRISPR systems in plants. Advanced CRISPR functionalities and delivery options demonstrated in plants are primarily reviewed but new technologies just coming to the forefront of CRISPR development, or those on the horizon, are briefly discussed. Topics covered are focused on the expansion of expression and delivery capabilities for CRISPR-Cas9 components and broadening targeting range through orthogonal Cas9 and Cpf1 proteins.

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Section: Introductionmentioning

confidence: 99%

Rapid Evolution of Manifold CRISPR Systems for Plant Genome Editing

Lowder

Malzahn

2016

Front. Plant Sci.

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“…Both QTLs are widely used in indica varieties, but are rarely exploited in japonica varieties. We designed and constructed a CRISPR/Cas9 vector that targets the disruption of both GS3 and Gn1a (Figures A, S1) using a previously‐described method (Shan et al ; Wang et al ). Five widely‐cultivated japonica varieties, namely Nanjing 9108 (N9108), Wuyunjing 27 (W27), Yangjing 4227 (Y4227), Zhejing 22 (Z22), and Zhejing 88 (Z88), were chosen for targeted mutation.…”

mentioning

confidence: 99%

QTL editing confers opposing yield performance in different rice varieties

Shen

Chün

et al. 2016

JIPB

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129

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Grain yield is one of the most important and complex trait for genetic improvement in crops; it is known to be controlled by a number of genes known as quantitative trait loci (QTLs). In the past decade, many yield-contributing QTLs have been identified in crops. However, it remains unclear whether those QTLs confer the same yield performance in different genetic backgrounds. Here, we performed CRISPR/Cas9-mediated QTL editing in five widely-cultivated rice varieties and revealed that the same QTL can have diverse, even opposing, effects on grain yield in different genetic backgrounds.

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“…To further confirm that the mutation of OsSPL resulted in the sterile phenotype, we employed the CRISPR‐Cas9 system to generate specific mutations at the OsSPL locus (Fig. S6a; Wang et al ., ). We obtained 34 T 0 transgenic rice plants, two of which had 1‐bp and 2‐bp homozygous deletions at the target site (Fig.…”

Section: Resultsmentioning

confidence: 97%

“…The target sequence was synthesized and then fused with Aar I linearized SK-gRNA vector. The intermediate vector was introduced into CRISPR-Cas9 binary vector pC1300-Cas9 (Wang et al, 2015). The construct was used for Agrobacterium-mediated transformation in japonica rice cultivar Yandao 8.…”

Section: Crispr-cas9 Targeting Of Ossplmentioning

confidence: 99%

OsSPL regulates meiotic fate acquisition in rice

et al. 2018

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In angiosperms, the key step in sexual reproduction is successful acquisition of meiotic fate. However, the molecular mechanism determining meiotic fate remains largely unknown. Here, we report that OsSPOROCYTELESS (OsSPL) is critical for meiotic entry in rice (Oryza sativa). We performed a large-scale genetic screen of rice sterile mutants aimed to identify genes regulating meiotic entry and identified OsSPL using map-based cloning. We showed that meiosis-specific callose deposition, chromatin organization, and centromere-specific histone H3 loading were altered in the cells corresponding to pollen mother cells in Osspl anthers. Global transcriptome analysis showed that the enriched differentially expressed genes in Osspl were mainly related to redox status, meiotic process, and parietal cell development. OsSPL might form homodimers and interact with TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) transcription factor OsTCP5 via the SPL dimerization and TCP interaction domain. OsSPL also interacts with TPL (TOPLESS) corepressors, OsTPL2 and OsTPL3, via the EAR motif. Our results suggest that the OsSPL-mediated signaling pathway plays a crucial role in rice meiotic entry, which appears to be a conserved regulatory mechanism for meiotic fate acquisition in angiosperms.

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A Simple CRISPR/Cas9 System for Multiplex Genome Editing in Rice

Cited by 121 publications

References 11 publications

Rapid Evolution of Manifold CRISPR Systems for Plant Genome Editing

Rapid Evolution of Manifold CRISPR Systems for Plant Genome Editing

QTL editing confers opposing yield performance in different rice varieties

OsSPL regulates meiotic fate acquisition in rice

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