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CRISPR-Cas12a-Based DNA Detection for Fast Pathogen Diagnosis and GMO Test in Plants

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CRISPR-Cas Methods

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Abstract

DNA test is widely used in plant pathogen diagnosis and genetically modified organism (GMO) administration. To date, a low-cost, user-friendly, and field-deployable DNA test method with high accuracy and sensitivity is still limited. Recently, the RNA programmable nuclease of CRISPR-Cas is engineered as a new nucleic acid detection platform, providing a novel and promising DNA test strategy for in-field crop disease diagnosis and GMO identification. In this study, we describe an all-paper-based DNA test method using CRISPR-Cas12a. This method combines filter-paper-based DNA purification, recombinase polymerase amplification, target gene detection with Cas12a, and lateral flow assay. Owing to its simplicity, efficiency, robustness, and low-cost, this all-paper-based Cas12a DNA test method could be easily applied in field for crop disease diagnosis and GMO test.

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Acknowledgments

We thank Profs. Yongjun Lin and Hao Chen for providing the Bt-rice plants. This study was supported by the National Natural Science Foundation of China (31622047), National Transgenic Science and Technology program (2018ZX08010-05B), National Key Laboratory of Crop Genetic Improvement, and State Key Laboratory of Hybrid Rice.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement and Hubei Key Laboratory of Plant Pathology, Huazhong Agricultural University, Wuhan, China

    Yun-Mu Zhang & Kabin Xie

  2. Department of Plant Pathology and Environmental Microbiology, The Pennsylvania State University, University Park, PA, USA

    Yinong Yang

Authors
  1. Yun-Mu Zhang
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  2. Yinong Yang
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  3. Kabin Xie
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Corresponding author

Correspondence to Kabin Xie .

Editor information

Editors and Affiliations

  1. Institute of Biotechnology and Genetic Engineering (IBGE), Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh

    M. Tofazzal Islam

  2. ICAR-National Rice Research Institute, Cuttack, Odisha, India

    Kutubuddin Ali Molla

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Zhang, YM., Yang, Y., Xie, K. (2021). CRISPR-Cas12a-Based DNA Detection for Fast Pathogen Diagnosis and GMO Test in Plants. In: Islam, M.T., Molla, K.A. (eds) CRISPR-Cas Methods. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1657-4_15

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  • DOI: https://doi.org/10.1007/978-1-0716-1657-4_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1656-7

  • Online ISBN: 978-1-0716-1657-4

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