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Silencing of Stress-Regulated miRNAs in Plants by Short Tandem Target Mimic (STTM) Approach

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Plant Stress Tolerance

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1631))

Abstract

In plants, microRNAs (miRNAs) regulate more than hundred target genes comprising largely transcription factors that control growth and development as well as stress responses. However, the exact functions of miRNA families could not be deciphered because each miRNA family has multiple loci in the genome, thus are functionally redundant. Therefore, an ideal approach to study the function of a miRNA family is to silence the expression of all members simultaneously, which is a daunting task. However, this can be partly overcome by Target Mimic (TM) approach that can knockdown an entire miRNA family. STTM is a modification of TM approach and complements it. STTMs have been successfully used in monocots and dicots to block miRNA functions. miR159 has been shown to be differentially regulated by various abiotic stresses including ABA in various plant species. Here, we describe in detail the protocol for designing STTM construct to block miR159 functions in Arabidopsis, with the potential to apply this technique on a number of other stress-regulated miRNAs in plants.

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Acknowledgements

This study was partially supported by funds from Henan Agricultural University (HAU) and NSFC (31571679), China. G.T. is supported by the National Science Foundation (NSF; grants IOS-1048216 and IOS-1340001). S.T. is supported by a postdoctoral scholarship from HAU.

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Authors and Affiliations

  1. College of Agronomy, Henan Agricultural University, Zhengzhou, 450002, China

    Sachin Teotia & Guiliang Tang

  2. School of Biotechnology, Gautam Buddha University, Greater Noida, 201312, India

    Sachin Teotia

  3. Department of Biological Sciences, Michigan Technological University, 1400 Townsend Dr, Houghton, MI, 49931, USA

    Sachin Teotia & Guiliang Tang

Authors
  1. Sachin Teotia
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  2. Guiliang Tang
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Corresponding author

Correspondence to Guiliang Tang .

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Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, Oklahoma, USA

    Ramanjulu Sunkar

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Teotia, S., Tang, G. (2017). Silencing of Stress-Regulated miRNAs in Plants by Short Tandem Target Mimic (STTM) Approach. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 1631. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7136-7_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7136-7_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7134-3

  • Online ISBN: 978-1-4939-7136-7

  • eBook Packages: Springer Protocols

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