Dataset Information

Atypical Splicing Accompanied by Skipping Conserved Micro-exons Produces Unique WRINKLED1, An AP2 Domain Transcription Factor in Rice Plants.

ABSTRACT: WRINKLED1 (WRI1), an AP2 domain transcription factor, is a master regulator of oil synthesis in plant seeds. Its closely related proteins (WRIs) are also involved in regulating the synthesis of fatty acids, which play a role in producing oils, membranes, and other important components in plants. We found two WRI1 genes, OsWRI1-1 and OsWRI1-2, and two additional WRI1 homologs, OsWRI3 and OsWRI4, in the rice genome. OsWRI1 was ubiquitously expressed in rice plants, including developing seeds. However, OsWRI3 was only significantly expressed in the leaf blade and OsWRI4 was not expressed at all. OsWRI1-1 contains amino acid sequence GCL instead of VYL, which is encoded by an independent 9-bp micro-exon that is conserved in many plant species. We found that the GCL sequence was produced by an atypical splicing accompanied by skipping of the micro-exon. Furthermore, OsWRI1-1 highly activates the transcription of the promoter for the biotin carboxyl transferase 2 gene in Arabidopsis, but its activity was reduced by amino acid replacement or deletion of the GCL sequence in a transient assay using Arabidopsis cells. Our results indicated that atypical splicing produced unique WRI1 in rice plants.

SUBMITTER: Mano F

PROVIDER: S-EPMC6681275 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Atypical Splicing Accompanied by Skipping Conserved Micro-exons Produces Unique WRINKLED1, An AP2 Domain Transcription Factor in Rice Plants.

Mano Fumiya F Aoyanagi Takuya T Kozaki Akiko A

Plants (Basel, Switzerland) 20190704 7

WRINKLED1 (WRI1), an AP2 domain transcription factor, is a master regulator of oil synthesis in plant seeds. Its closely related proteins (WRIs) are also involved in regulating the synthesis of fatty acids, which play a role in producing oils, membranes, and other important components in plants. We found two WRI1 genes, OsWRI1-1 and OsWRI1-2, and two additional WRI1 homologs, OsWRI3 and OsWRI4, in the rice genome. OsWRI1 was ubiquitously expressed ...[more]

PMID: 31277505

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Project description:Antisense oligonucleotide-mediated exon skipping is a promising therapeutic approach for the treatment of various genetic diseases and a therapy which has gained significant traction in recent years following FDA approval of new antisense-based drugs. Exon skipping for Duchenne muscular dystrophy (DMD) works by modulating dystrophin pre-mRNA splicing, preventing incorporation of frame-disrupting exons into the final mRNA product while maintaining the open reading frame, to produce a shortened-yet-functional protein as seen in milder Becker muscular dystrophy (BMD) patients. Exons 45-55 skipping in dystrophin is potentially applicable to approximately 47% of DMD patients because many mutations occur within this "mutation hotspot." In addition, patients naturally harboring a dystrophin exons 45-55 in-frame deletion mutation have an asymptomatic or exceptionally mild phenotype compared to shorter in-frame deletion mutations in this region. As such, exons 45-55 skipping could transform the DMD phenotype into an asymptomatic or very mild BMD phenotype and rescue nearly a half of DMD patients. In addition, this strategy is potentially applicable to some BMD patients as well, who have in-frame deletion mutations in this region. As the degree of exon skipping correlates with therapeutic outcomes, reliable measurements of exon skipping efficiencies are essential to the development of novel antisense-mediated exon skipping therapeutics. In the case of DMD, researchers have often relied upon human muscle fibers obtained from muscle biopsies for testing; however, this method is highly invasive and patient myofibers can display limited proliferative ability. To overcome these challenges, researchers can generate myofibers from patient fibroblast cells by transducing the cells with a viral vector containing MyoD, a myogenic regulatory factor. Here, we describe a methodology for assessing dystrophin exons 45-55 multiple skipping efficiency using antisense oligonucleotides in human muscle cells derived from DMD patient fibroblast cells.

Dataset Information

Atypical Splicing Accompanied by Skipping Conserved Micro-exons Produces Unique WRINKLED1, An AP2 Domain Transcription Factor in Rice Plants.

Publications

Atypical Splicing Accompanied by Skipping Conserved Micro-exons Produces Unique WRINKLED1, An AP2 Domain Transcription Factor in Rice Plants.

Similar Datasets

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