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Related Experiment Video

Updated: Mar 15, 2026

mRNA Interactome Capture from Plant Protoplasts
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Live Cell Microscopy-Based RNAi Screening in the Moss Physcomitrella patens.

Tomohiro Miki1, Yuki Nakaoka1, Gohta Goshima2

  • 1Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|September 2, 2016
PubMed
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Proceedings of the National Academy of Sciences of the United States of America·2024

This study presents a microscopy-based RNA interference (RNAi) screening method for moss cells. The technique efficiently identifies genes involved in plant cell growth and division.

Area of Science:

  • Plant Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • RNA interference (RNAi) is crucial for identifying genes in cellular processes.
  • Physcomitrella patens is an emerging model organism for plant cell biology research.
  • Efficient screening methods are needed to study gene function in P. patens.

Purpose of the Study:

  • To establish a microscopy-based RNAi screening protocol for Physcomitrella patens protonemal cells.
  • To enable efficient and effective screening of over 100 genes.
  • To facilitate the study of genes involved in cell division, organelle distribution, and cell growth.

Main Methods:

  • Utilizing conditional (inducible) RNAi vectors for controlled gene silencing.
  • Developing transgenic moss lines with integrated RNAi vectors.
Keywords:
Inducible RNAiKinesinLive cell imagingPhyscomitrella patensProtonemal cell

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  • Employing time-lapse fluorescent microscopy for dynamic cellular process observation.

    • Main Results:

    • The developed method allows for effective and efficient RNAi screening in P. patens.
    • Over 100 genes involved in key cellular processes were screened.
    • The protocol is suitable for studying mitotic cell division, organelle distribution, and cell growth.

    Conclusions:

    • This protocol provides a robust platform for functional genomics in Physcomitrella patens.
    • The method enhances the study of plant cell biology using RNA interference.
    • It facilitates large-scale gene function analysis in a model moss system.