Acquisition of seed dormancy breaking in rice (Oryza sativa L.) via CRISPR/Cas9-targeted mutagenesis of OsVP1 gene

Jung, Yu Jin; Lee, Hyo Ju; Bae, Sangsu; Kim, Jong Hee; Kim, Dong Hyen; Kim, Hee Kyoung; Nam, Ki Hong; Nogoy, Franz Marielle; Cho, Yong Gu; Kang, Kwon Kyoo

doi:10.1007/s11816-019-00580-x

Cited by 25 publications

(22 citation statements)

References 39 publications

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“…The remaining lines were mostly single base deletions and insertions ( Figure 3 ). Jung [ 33 ] reported the deletion of a target gene in tomatoes via CRISPR/Cas9, and the frequency of deletion correlated with the target size. For example, it was about 67% for 1 to 10 bp deletion, but only 3% for 10 bp or higher deletion.…”

Section: Discussionmentioning

confidence: 99%

“…The T7E1 assay was performed in the same way as previously reported by Jung [ 33 ]. In the pBAtC vector, the Cas9 gene was regulated by the 35S mosaic virus promoter, and the sgRNA was controlled by the Arabidopsis U6 promoter [ 33 ]. DNA oligos corresponding to the designed sgRNAs were synthesized by Bioneer co., Ltd. (Dajeon, Korea) ( Table S7 ) and the dimer was cloned into plant expression vector, pBAtC.…”

Section: Methodsmentioning

confidence: 99%

“…First, to identify transformants from regenerated plants, PCR analysis was performed using NPTII gene-specific primers. Then, targeted deep sequence analysis was performed as described by Jung [ 33 ]. A list of all primers used for targeted deep sequencing was shown in Table S2 .…”

Section: Methodsmentioning

confidence: 99%

“…Recent genome editing techniques have shown the possibility of inducing gene mutations at desired genomic DNA locations using various types of site-specific nucleases [ 31 ]. Among them, the CRISPR/Cas9 system has been known to be a powerful genome editing tool in regards to plants and many other organisms [ 32 , 33 ]. The advantage of these systems is the precise and efficient introduction of mutations at the target site.…”

Section: Introductionmentioning

confidence: 99%

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Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato

Jung

Kim

Lee

et al. 2020

Plants

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The utilization of male sterility into hybrid seed production reduces its cost and ensures high purity of tomato varieties because it does not produce pollen and has exserted stigmas. Here, we report on the generation of gene edited lines into male sterility phenotype by knockout of SlMS10 gene (Solyc02g079810) encoding the bHLH transcription factor that regulates meiosis and cell death of the tapetum during microsporogenesis in the tomato. Twenty-eight gene edited lines out of 60 transgenic plants were selected. Of these, eleven different mutation types at the target site of the SlMS10 gene were selected through deep sequencing analysis. These mutations were confirmed to be transmitted to subsequent generations. The null lines without the transferred DNA (T-DNA) were obtained by segregation in the T1 and T2 generations. In addition, we showed that the cr-ms10-1-4 mutant line exhibited dysfunctional meiosis and abnormal tapetum during flower development, resulting in no pollen production. RT-PCR analysis showed that the most genes associated with pollen and tapetum development in tomatoes had lower expression in the cr-ms10-1-4 mutant line compared to wild type. We demonstrate that modification of the SlMS10 gene via CRISPR/Cas9-mediated genome editing results in male sterility of tomato plants. Our results suggest an alternative approach to generating male sterility in crops.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato

Jung

Kim

Lee

et al. 2020

Plants

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“…Jung et al (2019a, b) showed CRISPR/Cas9-mediated gene mutagenesis in rice. Jung et al (2019a) performed the CRISPR/Cas9-mediated editing of viviparous-1 (OsVP1) gene to break seed dormancy in rice and obtained transgenefree lines in the T 1 generation that exhibited faster germination rate without compromising the other agronomic traits compared with wild-type plants under field conditions. Jung et al (2019b) performed targeted mutagenesis of F3′H, DFR and LDOX genes related to anthocyanin biosynthesis in black rice (Oryza sativa L.).…”

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confidence: 99%