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Related Experiment Videos

Gene expression and methylation patterns in cloned embryos.

Christine Wrenzycki1, Doris Herrmann, Claudia Gebert

  • 1Department of Biotechnology, Institute for Animal Science, FAL, Mariensee, Neustadt, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|September 22, 2006
PubMed
Summary

Large Offspring Syndrome (LOS) in cloned and in vitro-produced embryos is linked to abnormal gene expression, potentially due to DNA methylation changes. Understanding these epigenetic alterations is key to improving embryo development.

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Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Epigenetics

Background:

  • Offspring from nuclear transfer (NT) and in vitro production (IVP) often exhibit abnormalities like high birthweight, termed Large Offspring Syndrome (LOS).
  • LOS is hypothesized to stem from persistent gene expression aberrations, beginning in preimplantation stages, with epigenetic modifications, particularly DNA methylation, implicated.

Purpose of the Study:

  • To investigate the molecular mechanisms underlying Large Offspring Syndrome (LOS) in mammalian embryos.
  • To compare gene expression patterns in NT- and IVP-derived embryos with in vivo-derived embryos to establish a physiological standard.

Main Methods:

  • Utilized sensitive reverse transcription polymerase chain reaction (RT-PCR) assays to analyze messenger RNA (mRNA) expression levels in single embryos.

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  • Employed bisulfite sequencing to assess the DNA methylation status of specific genes.

    • Main Results:

    • Observed numerous mRNA expression aberrations in IVP and NT-derived embryos, including gene suppression, de novo overexpression, and significant upregulation or downregulation.
    • These findings highlight deviations from the normal gene expression patterns seen in in vivo-derived embryos.

    Conclusions:

    • Aberrant DNA methylation and subsequent gene expression changes are likely contributors to Large Offspring Syndrome (LOS).
    • Further research into these molecular mechanisms is crucial for enhancing the viability of embryos produced through assisted reproductive technologies and improving mammalian biotechnologies.