Comparison between the Repression Potency of siRNA Targeting the Coding Region and the 3′-Untranslated Region of mRNA
| Autor: | Chih-Ying Chen, Lo-Chun Au, Ching-Fang Lai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Genetics
Small interfering RNA General Immunology and Microbiology Article Subject Three prime untranslated region Trans-acting siRNA lcsh:R Fireflies lcsh:Medicine General Medicine Biology General Biochemistry Genetics and Molecular Biology Stop codon Open reading frame Open Reading Frames Luciferases Firefly Gene silencing Coding region Animals RNA Interference RNA Messenger RNA Small Interfering Psychological repression 3' Untranslated Regions Research Article |
| Zdroj: | BioMed Research International, Vol 2013 (2013) BioMed Research International |
| ISSN: | 2314-6141 2314-6133 |
| Popis: | Small interfering RNAs (siRNAs) are applied for post-transcriptional gene silencing by binding target mRNA. A target coding region is usually chosen, although the3′-untranslated region (3′-UTR) can also be a target. This study elucidates whether the coding region or3′-UTR elicits higher repression. pFLuc and pRLuc are two reporter plasmids. A segment ofFLucgene was PCR-amplified and inserted behind the stop codon of theRLucgene of the pRLuc. Similarly, a segment ofRLucgene was inserted behind the stop codon ofFLuc. Two siFLuc and two siRLuc were siRNAs designed to target the central portions of these segments. Therefore, the siRNA encountered the same targets and flanking sequences. Results showed that the two siFLuc elicited higher repression when theFLucsegment resided in the coding region. Conversely, the two siRLuc showed higher repression when theRLucsegment was in the3′-UTR. These results indicate that both the coding region and the3′-UTR can be more effective targets. The thermodynamic stability of the secondary structures was analyzed. The siRNA elicited higher repression in the coding region when the target configuration was stable, and needed to be solved by translation. A siRNA may otherwise favor the target at3′-UTR. |
| Databáze: | OpenAIRE |
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