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Identification, stability and expression of Sirt1 antisense long non-coding RNA
Yu Wang1, Wei-Jun Pang1, Ning Wei1
1College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Gene
|February 1, 2014
Summary
Researchers discovered a novel natural antisense transcript (NAT) for Sirtuin 1 (Sirt1), a key protein in muscle development. This Sirt1 antisense long non-coding RNA (AS lncRNA) regulates Sirt1 expression during myogenesis.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Natural antisense transcripts (NATs) are crucial regulators of gene expression.
- Sirtuin 1 (Sirt1), a NAD-dependent deacetylase, plays a role in myogenesis.
- The existence and function of Sirt1 NATs during C2C12 cell differentiation were previously unknown.
Purpose of the Study:
- To identify and characterize a NAT for Sirtuin 1 (Sirt1).
- To investigate the role of this Sirt1 NAT in C2C12 myogenic differentiation.
- To elucidate the regulatory mechanism of Sirt1 AS lncRNA on Sirt1 expression.
Main Methods:
- Sequencing and bioinformatic analysis to identify Sirt1 NAT.
- Quantitative analysis of Sirt1 mRNA and Sirt1 AS lncRNA expression during C2C12 differentiation.
- Actinomycin D treatment to determine RNA half-life.
- Overexpression studies of Sirt1 AS lncRNA and its mutant in C2C12 cells.
- Investigation of the interaction between Sirt1 AS lncRNA, miR-34a, and Sirt1 mRNA.
Main Results:
- A novel Sirt1 antisense long non-coding RNA (AS lncRNA) was identified.
- Sirt1 AS lncRNA expression was higher in spleen and lower in muscle tissue.
- Both Sirt1 mRNA and Sirt1 AS lncRNA decreased during C2C12 differentiation, while miR-34a increased.
- Sirt1 AS lncRNA demonstrated greater stability (10h half-life) compared to Sirt1 mRNA (6h).
- Overexpression of Sirt1 AS lncRNA increased Sirt1 protein levels, while a mutant form had no effect.
- Sirt1 AS lncRNA counteracted miR-34a-mediated downregulation of Sirt1 mRNA.
Conclusions:
- A novel Sirt1 NAT, Sirt1 AS lncRNA, was identified.
- Sirt1 AS lncRNA is implicated in myogenesis by regulating Sirt1 expression.
- Sirt1 AS lncRNA acts as a more stable regulator of Sirt1 compared to Sirt1 mRNA.
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