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Single Oocyte Bisulfite Mutagenesis
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Transcriptome Profiling of Single Mouse Oocytes.

Maud Borensztein1, Laurène Syx2, Nicolas Servant3

  • 1Institut Curie, PSL Research University, CNRS UMR3215, INSERM U934, UPMC Paris-Sorbonne, Paris, 75005, France. maud.borensztein@curie.fr.

Methods in Molecular Biology (Clifton, N.J.)
|July 2, 2018
PubMed
Summary
This summary is machine-generated.

This study details a single-cell RNA sequencing (scRNAseq) protocol for analyzing mouse oocyte transcriptomes. This method allows for gene expression analysis from limited samples, crucial for studying mutant mouse models.

Keywords:
Gene expressionMaternal poolSingle-cell RNA-sequencingSingle-cell bioinformatics pipeline

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Area of Science:

  • Developmental Biology
  • Genomics
  • Molecular Biology

Background:

  • Limited starting material from oocytes and embryos poses challenges for transcriptomic analysis.
  • Single-cell RNA sequencing (scRNAseq) offers a solution for studying gene expression at the individual cell level.
  • scRNAseq is particularly valuable when working with mutant mouse models where sample quantity is restricted.

Purpose of the Study:

  • To outline a detailed single-cell RNA sequencing (scRNAseq) protocol specifically for mouse oocyte transcriptomes.
  • To provide a comprehensive method for analyzing gene expression in individual oocytes.
  • To facilitate research on oocyte biology, especially in the context of genetic variations.

Main Methods:

  • Single-cell isolation from mouse oocytes.
  • Complementary DNA (cDNA) amplification from the isolated single cells.
  • High-throughput sequencing of amplified cDNA.
  • Bioinformatics pipeline for genome-wide gene expression analysis and comparison.

Main Results:

  • The protocol successfully enables the detection and quantification of mature RNAs in individual mouse oocytes.
  • The described bioinformatics pipeline allows for effective analysis and comparison of genome-wide gene expression patterns between oocytes.
  • The method is robust for studying transcriptomes from limited biological samples, such as those from mutant mice.

Conclusions:

  • The presented scRNAseq protocol is effective for studying mouse oocyte transcriptomes.
  • This method overcomes limitations associated with small sample sizes in oocyte and embryo research.
  • The protocol supports detailed analysis of gene expression variability at the single-cell level in oocytes.