Targeted mutation of GA20ox-2 gene using CRISPR/Cas9 system generated semi-dwarf phenotype in rice
| dc.contributor.author | Santoso, T J ...[at al] | |
| dc.contributor.other | Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian | en_US |
| dc.date.accessioned | 2021-03-17T05:41:45Z | |
| dc.date.available | 2021-03-17T05:41:45Z | |
| dc.date.issued | 2020 | |
| dc.description.abstract | Recently, the engineered CRISPR/Cas9 system has been applied to rapidly and efficiently modify the targeted gene(s) in a wide variety of plants. Recent studies of successful targeted mutagenesis using the CRISPR/Cas9 system with a single gRNA expression in rice plants have been reported. GA20ox-2 is a gene encoding an oxidase enzyme involved in the biosynthesis of gibberellin and linked to sd1 locus. A previous study revealed that mutation of this gene resulted in shorter stature of rice plant due to defects in the gibberellin’s signalling pathway. Here, we studied targeted mutation of OsGA20ox-2 gene in rice using the CRISPR/Cas9 system with the expression of two gRNAs. In this study, we introduced a single plasmid vector of CRISPR/Cas9 system harboring dual gRNAs to modify OsGA20ox-2 gene in a rice model cv. Kitaake via Agrobacterium-mediated transformation. Targeted mutagenesis of OsGA20ox-2 gene using CRISPR/Cas9 generated nine mutated rice lines with a mutation frequency of 90%. Most mutated lines (50%) had mutations in both OsGA20ox-2 gRNA. They resulted in homo-diallelic mutation type with 44 bp deletion, while three lines were heterozygous, one line was homo-diallelic with 2 bp insertion, and one line had no mutation. The K15 mutated rice line was identified as a homozygous two-nucleotide insertion and had the semi-dwarf phenotype, demonstrating that OsGA20ox-2 gene had been disrupted. | en_US |
| dc.identifier.isbn | - | |
| dc.identifier.uri | https://repository.pertanian.go.id/handle/123456789/12063 | |
| dc.language.iso | en | en_US |
| dc.publisher | IAARD Press | en_US |
| dc.subject | rice (Oryza sativa L.), CRISPR/Cas9 system, targeted mutagenesis, semi-dwarf. | en_US |
| dc.title | Targeted mutation of GA20ox-2 gene using CRISPR/Cas9 system generated semi-dwarf phenotype in rice | en_US |
| dc.type | Article | en_US |