An efficient and rapid method to detect and verify natural antisense transcripts of animal genes
| Autor: | Bi-xiao Li, Li Zhang, Qinghua Nie, Xiquan Zhang, Jing-e Ma, Shudai Lin, Mei Xiao, An LiLong, Dexiang Zhang, Rui Zhao, Feng-Fang Qiu |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Agriculture (General) Plant Science Biology natural antisense transcripts Biochemistry S1-972 03 medical and health sciences Food Animals Sense (molecular biology) Genetics Ecology long non-coding RNA detection method RNA Nuclease protection assay RNA sequencing Non-coding RNA Long non-coding RNA Antisense RNA RNA silencing 030104 developmental biology Animal Science and Zoology transcription orientation Tsix Agronomy and Crop Science Food Science |
| Zdroj: | Journal of Integrative Agriculture, Vol 15, Iss 9, Pp 2070-2076 (2016) |
| ISSN: | 2095-3119 |
| Popis: | High-throughput sequencing has identified a large number of sense-antisense transcriptional pairs, which indicates that these genes were transcribed from both directions. Recent reports have demonstrated that many antisense RNAs, especially lncRNA (long non-coding RNA), can interact with the sense RNA by forming an RNA duplex. Many methods, such as RNA-sequencing, Northern blotting, RNase protection assays and strand-specific PCR, can be used to detect the antisense transcript and gene transcriptional orientation. However, the applications of these methods have been constrained, to some extent, because of the high cost, difficult operation or inaccuracy, especially regarding the analysis of substantial amounts of data. Thus, we developed an easy method to detect and validate these complicated RNAs. We primarily took advantage of the strand specificity of RT-PCR and the single-strand specificity of S1 endonuclease to analyze sense and antisense transcripts. Four known genes, including mouse β-actin and Tsix (Xist antisense RNA), chicken LXN (latexin) and GFM1 (G elongation factor, mitochondrial 1), were used to establish the method. These four genes were well studied and transcribed from positive strand, negative strand or both strands of DNA, respectively, which represented all possible cases. The results indicated that the method can easily distinguish sense, antisense and sense-antisense transcriptional pairs. In addition, it can be used to verify the results of high-throughput sequencing, as well as to analyze the regulatory mechanisms between RNAs. This method can improve the accuracy of detection and can be mainly used in analyzing single gene and was low cost. |
| Databáze: | OpenAIRE |
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