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Related Concept Videos

MicroRNAs01:22

MicroRNAs

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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MicroRNA Expression Profiling, Target Identification, and Validation in Chondrocytes.

Tomoki Chiba1, Ryota Kurimoto1, Takahide Matsushima1

  • 1Department of Systems BioMedicine, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|December 14, 2020
PubMed
Summary

MicroRNAs (miRNAs) are small RNAs crucial for gene regulation in processes like bone formation and arthritis. This study details methods to analyze miRNA expression and function in cartilage cells, aiding arthritis research.

Keywords:
ArthritisCartilageChondrocytesExpression profilingTarget validationmiRNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are noncoding small RNAs regulating gene expression.
  • They are implicated in musculoskeletal development and arthritis pathogenesis.
  • A single miRNA can influence hundreds of target genes.

Purpose of the Study:

  • To outline experimental protocols for studying miRNA expression and function in chondrocytes.
  • To investigate the role of miRNAs in cartilage and arthritis.

Main Methods:

  • miRNA expression profiling using Next Generation Sequencing and TaqMan Array.
  • miRNA target prediction via TargetScan.
  • Cell-based reporter assays for target screening.
  • HITS-CLIP for analyzing miRNA-target interactions.

Main Results:

  • The study provides a comprehensive set of protocols for miRNA research in cartilage.
  • These methods enable detailed analysis of miRNA expression and function.
  • The described techniques facilitate understanding of gene regulation networks.

Conclusions:

  • The outlined protocols are essential for advancing research on miRNAs in cartilage and chondrocytes.
  • These methods will aid in understanding miRNA roles in arthritis pathogenesis.
  • This work provides a foundation for future miRNA-based therapeutic strategies in joint diseases.