Live Cell Microscopy-Based RNAi Screening in the Moss Physcomitrella patens

RNA interference (RNAi) is a powerful technique enabling the identification of the genes involved in a certain cellular process. Here, we discuss protocols for microscopy-based RNAi screening in protonemal cells of the moss Physcomitrella patens, an emerging model system for plant cell biology. Our...

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Bibliographic Details
Published in:	Methods in molecular biology (Clifton, N.J.) Vol. 1470; p. 225
Main Authors:	Miki, Tomohiro, Nakaoka, Yuki, Goshima, Gohta
Format:	Journal Article
Language:	English
Published:	United States 01-01-2016
Subjects:	Bryopsida - cytology Bryopsida - genetics Genetic Vectors High-Throughput Screening Assays - methods Histones - genetics Histones - metabolism Microscopy, Fluorescence - instrumentation Microscopy, Fluorescence - methods Mitosis Plant Cells Plant Proteins - genetics Plants, Genetically Modified RNA Interference Inducible RNAi Protonemal cell Physcomitrella patens Kinesin Live cell imaging
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Summary:	RNA interference (RNAi) is a powerful technique enabling the identification of the genes involved in a certain cellular process. Here, we discuss protocols for microscopy-based RNAi screening in protonemal cells of the moss Physcomitrella patens, an emerging model system for plant cell biology. Our method is characterized by the use of conditional (inducible) RNAi vectors, transgenic moss lines in which the RNAi vector is integrated, and time-lapse fluorescent microscopy. This method allows for effective and efficient screening of >100 genes involved in various cellular processes such as mitotic cell division, organelle distribution, or cell growth.
ISSN:	1940-6029
DOI:	10.1007/978-1-4939-6337-9_18