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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 the pre-miRNA...
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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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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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A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
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microRNA Expression Profiling: Technologies, Insights, and Prospects.

Christine Roden1, Stephen Mastriano1, Nayi Wang2

  • 1Department of Genetics, Yale Stem Cell Center and Yale Cancer Center, Yale University School of Medicine, 10 Amistad Street, Rm 237C, New Haven, CT, 06520-8005, USA.

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MicroRNA (miRNA) expression profiling is crucial for understanding miRNA functions and diagnostics. Careful data normalization and analysis of miRNA isoforms are essential for accurate interpretation and discovering new insights, such as miRNA

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • MicroRNA (miRNA) expression profiling is vital for elucidating miRNA biological functions and developing diagnostic tools.
  • Various technologies, including microarrays, next-generation sequencing, and RT-PCR, enable accurate miRNA quantification.

Purpose of the Study:

  • To discuss the challenges and best practices in miRNA expression profiling.
  • To highlight the impact of normalization and miRNA isoforms on data interpretation.
  • To present future prospects in single-cell miRNA variability and isoform function.

Main Methods:

  • Review of established miRNA expression profiling technologies (microarrays, NGS, bead-based, single-molecule, RT-PCR).
  • Discussion on experimental design and data analysis considerations, focusing on normalization and isoform detection.
  • Case study illustrating the impact of normalization on interpreting miRNA expression as a tumor suppressor.

Main Results:

  • Accurate miRNA quantification is achievable through multiple technologies.
  • Normalization strategies significantly influence data interpretation and biological conclusions.
  • Consideration of normalization revealed global miRNA expression as a potential tumor suppressor.

Conclusions:

  • Effective miRNA profiling requires meticulous experimental design and data analysis, particularly concerning normalization and isoforms.
  • Advanced miRNA profiling holds promise for understanding cellular heterogeneity and defining novel miRNA functions.
  • Future research directions include single-cell miRNA analysis and exploring the roles of miRNA isoforms.