論文 - 詳細
| RRC ID | 54880 |
|---|---|
| 著者 | Mitsuhashi H, Homma S, Beermann ML, Ishimaru S, Takeda H, Yu BK, Liu K, Duraiswamy S, Boyce FM, Miller JB. |
| タイトル | Efficient system for upstream mRNA trans-splicing to generate covalent, head-to-tail, protein multimers. |
| ジャーナル | Sci Rep |
| Abstract |
We present a plasmid-based system in which upstream trans-splicing efficiently generates mRNAs that encode head-to-tail protein multimers. In this system, trans-splicing occurs between one of two downstream splice donors in the sequence encoding a C-terminal V5 epitope tag and an upstream splice acceptor in the 5' region of the pCS2(+) host plasmid. Using deletion and fusion constructs of the DUX4 protein as an example, we found that this system produced trans-spliced mRNAs in which coding regions from independent transcripts were fused in phase such that covalent head-to-tail protein multimers were translated. For a cDNA of ~450 bp, about half of the expressed proteins were multimeric, with the efficiency of trans-splicing and extent of multimer expression decreasing as cDNA length increased. This system generated covalent heterodimeric proteins upon co-transfections of plasmids encoding separate proteins and did not require a long complementary binding domain to position mRNAs for trans-splicing. This plasmid-based trans-splicing system is adaptable to multiple gene delivery systems, and it presents new opportunities for investigating molecular mechanisms of trans-splicing, generating covalent protein multimers with novel functions within cells, and producing mRNAs encoding large proteins from split precursors. |
| 巻・号 | 9(1) |
| ページ | 2274 |
| 公開日 | 2019-2-19 |
| DOI | 10.1038/s41598-018-36684-7 |
| PII | 10.1038/s41598-018-36684-7 |
| PMID | 30783185 |
| PMC | PMC6381186 |
| MeSH | Genetic Engineering* HEK293 Cells HeLa Cells Homeodomain Proteins / genetics Homeodomain Proteins / metabolism Humans Plasmids / genetics* Plasmids / metabolism RNA, Messenger* / biosynthesis RNA, Messenger* / genetics Trans-Splicing* |
| IF | 4.011 |
| 引用数 | 0 |
| 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 | X(Twitter) |
| 各媒体での言及数の合計 | 1 |
| 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 | HeLa(RCB0007) |