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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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Transcriptome Profiling of In-Vivo Produced Bovine Pre-implantation Embryos Using Two-color Microarray Platform
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Profiling bovine blastocyst microRNAs using deep sequencing.

R Pasquariello1, B Fernandez-Fuertes2, F Strozzi3

  • 1Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territori, Università degli Studi di Milano, Via Celoria 2, 20133, Milan, Italy.

Reproduction, Fertility, and Development
|September 15, 2016
PubMed
Summary
This summary is machine-generated.

This study optimized microRNA (miRNA) extraction from bovine blastocysts, enabling reliable profiling of these crucial developmental molecules. The findings provide a foundation for understanding miRNA roles in early embryogenesis and reproductive health.

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Area of Science:

  • Reproductive Biology
  • Molecular Biology
  • Genomics

Background:

  • MicroRNAs (miRNAs) are critical regulators of reproductive functions, including oocyte maturation and embryonic development.
  • Profiling miRNAs in embryos is challenging due to low RNA content and high input requirements for library preparation.
  • Deep sequencing enables comprehensive miRNA analysis, but efficient extraction from limited embryonic samples is key.

Purpose of the Study:

  • To compare three RNA extraction methods for bovine blastocyst miRNA profiling.
  • To identify abundant and novel microRNAs in bovine blastocysts.
  • To investigate the functional relevance of identified miRNAs in early embryonic development.

Main Methods:

  • Comparison of three distinct RNA extraction procedures.
  • Library preparation and deep sequencing of pooled bovine blastocysts (30 per pool).
  • Bioinformatic analysis using miRDeep software for miRNA discovery and target prediction.

Main Results:

  • High concordance (14/15) in the most abundant miRNAs across all three extraction methods.
  • Identification of 1363 unique miRNA sequences, with bta-miR-10b and bta-miR-378 being the most abundant.
  • Identified target genes were significantly associated with cancer-related canonical pathways.

Conclusions:

  • Established reliable protocols for bovine blastocyst miRNA analysis, overcoming low RNA content challenges.
  • The identified miRNAs and their target pathways hold biological relevance for early embryogenesis.
  • Results demonstrate consistency across methods and species, supporting broad applicability.