title: Live Cell Microscopy-Based RNAi Screening in the Moss Physcomitrella patens.
abstract: 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.
authors: Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.
authors: Tomohiro
authors: T
authors: Miki
authors: Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.
authors: Yuki
authors: Y
authors: Nakaoka
authors: Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan. goshima@bio.nagoya-u.ac.jp.
authors: Gohta
authors: G
authors: Goshima
mesh_terms: D019068:Bryopsida / Q000166:cytology* / Q000235:genetics*; D005822:Genetic Vectors; D057166:High-Throughput Screening Assays / Q000379:methods*; D006657:Histones / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008938:Mitosis; D059828:Plant Cells; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D034622:RNA Interference
chemical_list: D006657:Histones; D010940:Plant Proteins
keywords: Inducible RNAi; Kinesin; Live cell imaging; Physcomitrella patens; Protonemal cell