Reference - Detail
| RRC ID | 51833 |
|---|---|
| Author | Kobayashi T, Yagi Y, Nakamura T. |
| Title | Development of Genome Engineering Tools from Plant-Specific PPR Proteins Using Animal Cultured Cells. |
| Journal | 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. |
| Volume | 1469 |
| Pages | 147-55 |
| Published | 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 |
| Times Cited | 1 |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | 293T(RCB2202) |