Here, we present a protocol for labeling and live visualization of RNA-protein complexes in the form of ribonucleoprotein particles (RNPs) in tobacco pollen tubes. We describe steps for constructing RNA-pp7/MS2 tag and biolistic gene-gun-mediated pollen transformation. We then provide detailed procedures for RNA labeling using PP7 aptamer nascent RNA tagging and a fluorescently labeled Pseudomonas aeruginosa PP7 bacteriophage coat protein (PCP) reporter that binds to PP7 RNA stem loops. This protocol is adaptable to other cell types by employing tissue-specific promoters.

Laboratory of Pollen Biology Institute of Experimental Botany of the Czech Academy of Sciences Rozvojová 263 165 00 Prague Czech Republic

Laboratory of Pollen Biology Institute of Experimental Botany of the Czech Academy of Sciences Rozvojová 263 165 00 Prague Czech Republic; Department of Experimental Plant Biology Faculty of Science Charles University Viničná 5 128 44 Prague Czech Republic

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