論文 - 詳細
| RRC ID | 46087 |
|---|---|
| 著者 | Minasaki R, Rudel D, Eckmann CR. |
| タイトル | Increased sensitivity and accuracy of a single-stranded DNA splint-mediated ligation assay (sPAT) reveals poly(A) tail length dynamics of developmentally regulated mRNAs. |
| ジャーナル | RNA Biol |
| Abstract |
Poly(A) tail length is a readout of an mRNA's translatability and stability, especially in developmental systems. PolyAdenylation Test (PAT) assays attempt to quickly measure the average poly(A) tail length of RNAs of experimental interest. Here we present sPAT, splint-mediated PAT, a procedure that uses a DNA splint to aid in the ligation of an RNA-tag to the poly(A) tail of an mRNA. In comparison to other PAT methodologies, including ePAT, sPAT is highly sensitive to low-abundance mRNAs, gives a more accurate profile of the poly(A) tail distribution, and requires little starting material. To demonstrate its strength, we calibrated sPAT on defined poly(A) tails of synthetic mRNAs, reassessed developmentally regulated poly(A) tail-length changes of known mRNAs from established model organisms, and extended it to the emerging evolutionary developmental nematode model Pristionchus pacificus. Lastly, we used sPAT to analyze the contribution of the two cytoplasmic poly(A) polymerases GLD-2 and GLD-4, and the deadenylase CCR-4, onto Caenorhabditis elegans gld-1 mRNA that encodes a translationally controlled tumor suppressor whose poly(A) tail length measurement proved elusive. |
| 巻・号 | 11(2) |
| ページ | 111-23 |
| 公開日 | 2014-1-1 |
| DOI | 10.4161/rna.27992 |
| PII | 27992 |
| PMID | 24526206 |
| PMC | PMC3973730 |
| MeSH | Animals DNA, Single-Stranded / metabolism* Evolution, Molecular Genetic Techniques* Models, Animal Phylogeny Poly A / metabolism* RNA Stability RNA, Messenger / metabolism* Reproducibility of Results |
| IF | 5.35 |
| 引用数 | 11 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| 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 | 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 | |
| 線虫 | tm1312 |