論文 - 詳細
| RRC ID | 51833 |
|---|---|
| 著者 | Kobayashi T, Yagi Y, Nakamura T. |
| タイトル | Development of Genome Engineering Tools from Plant-Specific PPR Proteins Using Animal Cultured Cells. |
| ジャーナル | Methods Mol Biol |
| Abstract |
The pentatricopeptide repeat (PPR) motif is a sequence-specific RNA/DNA-binding module. Elucidation of the RNA/DNA recognition mechanism has enabled engineering of PPR motifs as new RNA/DNA manipulation tools in living cells, including for genome editing. However, the biochemical characteristics of PPR proteins remain unknown, mostly due to the instability and/or unfolding propensities of PPR proteins in heterologous expression systems such as bacteria and yeast. To overcome this issue, we constructed reporter systems using animal cultured cells. The cell-based system has highly attractive features for PPR engineering: robust eukaryotic gene expression; availability of various vectors, reagents, and antibodies; highly efficient DNA delivery ratio (>80 %); and rapid, high-throughput data production. In this chapter, we introduce an example of such reporter systems: a PPR-based sequence-specific translational activation system. The cell-based reporter system can be applied to characterize plant genes of interested and to PPR engineering. |
| 巻・号 | 1469 |
| ページ | 147-55 |
| 公開日 | 2016-1-1 |
| DOI | 10.1007/978-1-4939-4931-1_11 |
| PMID | 27557692 |
| MeSH | 5' Untranslated Regions Animals Arabidopsis Proteins / genetics* Arabidopsis Proteins / metabolism Binding Sites Carrier Proteins / genetics* Carrier Proteins / metabolism Cells, Cultured Eukaryotic Initiation Factor-4G / genetics Eukaryotic Initiation Factor-4G / metabolism Genetic Engineering / methods* Genetic Vectors HEK293 Cells Humans Recombinant Fusion Proteins / genetics* Recombinant Fusion Proteins / metabolism Repetitive Sequences, Amino Acid Transfection |
| 引用数 | 1 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | 293T(RCB2202) |