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Published on: August 29, 2018
Identification and characterization of microRNAs from porcine skeletal muscle
Abstract:
MicroRNAs (miRNAs) are a class of non-coding RNAs that negatively regulate gene expression at the post-transcriptional level. There is increasing evidence to suggest that miRNAs participate in muscle development in mice and humans; however, few studies have focused on miRNAs in porcine muscle tissue. Here, we experimentally detected and identified conserved and unique miRNAs from porcine skeletal muscle. Fifty-seven distinct miRNAs were identified, of which 39 have not been reported earlier in the pig. Of these, two miRNAs appear to be novel and pig-specific. Surprisingly, these two differ only by a single nucleotide. A part of their primary transcript was cloned and confirmed by sequencing analysis. Alignment of the two sequences using ClustalW showed that the precursor sequences were almost identical, but the flanking sequences were different, indicating that these two novel miRNAs may represent rapidly evolving miRNAs in the pig genome. The expression patterns of eight miRNAs were characterized by real-time polymerase chain reaction of eight pig tissue samples. The ssc-let-7e and ssc-miR-181b miRNAs were expressed in all tissues analysed. The ssc-let-7c, ssc-miR-125b, ssc-miR-new1 and ssc-miR-new2 miRNAs were expressed in several tissues, while ssc-miR-122 and ssc-miR-206 were specifically expressed in the liver and muscle respectively. Our results add to existing data on porcine miRNAs and are useful for investigating the biological functions of miRNAs in porcine skeletal muscle development.
Insights
Researchers identified 57 distinct microRNAs (miRNAs) in porcine skeletal muscle, including two novel, pig-specific miRNAs. This discovery enhances understanding of miRNA roles in muscle development and evolution.
Area of Science:
- Genomics
- Molecular Biology
- Animal Science
Background:
- MicroRNAs (miRNAs) are crucial regulators of gene expression at the post-transcriptional level.
- Evidence suggests miRNA involvement in muscle development in mammals, but research in pigs is limited.
Purpose of the Study:
- To identify and characterize conserved and unique miRNAs in porcine skeletal muscle.
- To investigate the expression patterns of specific miRNAs across various pig tissues.
Main Methods:
- Experimental detection and identification of miRNAs using sequencing.
- Confirmation of novel miRNA transcripts via cloning and sequencing.
- Real-time polymerase chain reaction (PCR) for miRNA expression analysis across eight tissue types.
Main Results:
- Fifty-seven distinct miRNAs were identified in porcine skeletal muscle.
- Thirty-nine of these miRNAs were previously unreported in pigs.
- Two novel, potentially pig-specific miRNAs differing by a single nucleotide were discovered.
- Differential expression patterns were observed for eight characterized miRNAs, with some showing tissue-specific expression (e.g., ssc-miR-206 in muscle, ssc-miR-122 in liver).
Conclusions:
- This study expands the catalog of known porcine miRNAs.
- The identification of novel, rapidly evolving miRNAs provides insights into pig genome evolution.
- Characterized miRNA expression patterns are valuable for future research on porcine muscle development and function.
