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Genetic Suppressor Screen Using an Inducible FREE1-RNAi Line to Detect ESCRT Genetic Interactors in Arabidopsis thaliana

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The ESCRT Complexes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1998))

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Abstract

FREE1 (FYVE domain protein required for endosomal sorting 1), a newly identified component of endosomal sorting complex required for transport I (ESCRT I), plays multiple roles in regulating protein trafficking and organelle biogenesis in Arabidopsis. Similar to other ESCRT components, FREE1 is essential for plant growth and development because free1 mutant is seedling lethal. To identify key components that genetically interact with FREE1, we performed forward genetic suppressor screening using a dexamethasone (DEX)-inducible FREE1-RNAi line. Here we describe the detailed protocol of identifying novel FREE1 regulators using an inducible FREE1-RNAi line for the genetic suppressor screen. This protocol represents a whole procedure for identifying ESCRT genetic interactors in Arabidopsis thaliana.

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Acknowledgments

This work was supported by grants from the Research Grants Council of Hong Kong (CUHK466313 and 14130716 and CUHK2/CRF/11G, C4011-14R, C4012-16E, and AoE/M-05/12), the National Natural Science Foundation of China (31270226 and 31470294), and the Shenzhen Peacock Project (KQTD201101) (to L.J.). The authors declare no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory of Agrobiotechnology, Centre for Cell and Developmental Biology, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, China

    Qiong Zhao, Ying Zhu, Wenhan Cao, Jinbo Shen, Yong Cui, Shuxian Huang & Liwen Jiang

  2. CUHK Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, China

    Liwen Jiang

Authors
  1. Qiong Zhao
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  2. Ying Zhu
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  3. Wenhan Cao
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  4. Jinbo Shen
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  5. Yong Cui
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  6. Shuxian Huang
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  7. Liwen Jiang
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Editor information

Editors and Affiliations

  1. Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ.Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France

    Emmanuel Culetto

  2. Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ.Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France

    Renaud Legouis

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Zhao, Q. et al. (2019). Genetic Suppressor Screen Using an Inducible FREE1-RNAi Line to Detect ESCRT Genetic Interactors in Arabidopsis thaliana. In: Culetto, E., Legouis, R. (eds) The ESCRT Complexes. Methods in Molecular Biology, vol 1998. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9492-2_20

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  • DOI: https://doi.org/10.1007/978-1-4939-9492-2_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9491-5

  • Online ISBN: 978-1-4939-9492-2

  • eBook Packages: Springer Protocols

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