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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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Cryosectioning Method for Microdissection of Murine Colonic Mucosa
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MicroRNAs control intestinal epithelial differentiation, architecture, and barrier function.

Lindsay B McKenna1, Jonathan Schug, Anastassios Vourekas

  • 1Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19103, USA.

Gastroenterology
|July 28, 2010
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Summary

MicroRNAs (miRNAs) are crucial for intestinal health. Gene ablation of Dicer1, an miRNA-processing enzyme, disrupts intestinal epithelial differentiation and function, leading to impaired barrier function and inflammation.

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Area of Science:

  • Molecular Biology
  • Gastroenterology
  • Genetics

Background:

  • The role of microRNAs (miRNAs) in intestinal development and homeostasis remains largely unexplored.
  • Previous research has established miRNA importance in various tissue development.

Purpose of the Study:

  • To comprehensively profile miRNA expression in the mammalian intestinal mucosa.
  • To elucidate the functional contribution of miRNAs to intestinal homeostasis using genetic models.

Main Methods:

  • Ultrahigh throughput sequencing to determine the miRNA transcriptome of mouse intestinal mucosa.
  • High-throughput sequencing of RNA isolated by cross-linking immunoprecipitation (HITS-CLIP) to identify miRNA-mRNA targets.
  • Generation of intestinal epithelial-specific Dicer1-deficient mice using gene ablation.

Main Results:

  • Significant variation in miRNA abundance within the intestinal mucosa was observed, with mmu-miR-192 being the most abundant.
  • Dicer1-deficient mice exhibited disorganized intestinal epithelium, decreased goblet cells, increased crypt apoptosis, and accelerated cell migration.
  • Impaired intestinal barrier function, inflammation, and immune cell infiltration were noted in Dicer1-deficient mice.

Conclusions:

  • This study identified the complete repertoire of intestinal miRNAs.
  • Genetic ablation of Dicer1 demonstrated the essential role of miRNAs in intestinal epithelial differentiation and function.