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Knocking Out MicroRNA Genes in Rice with CRISPR-Cas9

  • Jianping Zhou
  • Zhaohui Zhong
  • Hongqiao Chen
  • Qian Li
  • Xuelian Zheng
  • Yiping Qi
  • Yong ZhangEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)

Abstract

Background: MicroRNAs (miRNAs) are small noncoding RNAs that play important roles in plant development and stress responses. Loss-of-function analysis of miRNA genes has been traditionally challenging due to lack of appropriate knockout tools. In this chapter, we describe a method of using CRISPR-Cas9 for knocking out microRNA genes in rice by Agrobacterium-mediated transformation. We also demonstrate single-strand conformation polymorphism (SSCP) as an effective genotyping method for screening CRISPR-Cas9-induced mutations.

Key words

MicroRNAs CRISPR-Cas9 Genome editing Rice SSCP 

Notes

Acknowledgments

This work was supported by grants including the National Science Foundation of China (31330017 and 31771486), the Sichuan Youth Science and Technology Foundation (2017JQ0005), the National Transgenic Major Project (2018ZX08022001-003), the Fundamental Research Funds for the Central Universities (ZYGX2016J119 and ZYGX2016J122) to YZ and XZ, and a start-up fund from the University of Maryland, College Park to YQ.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jianping Zhou
    • 1
  • Zhaohui Zhong
    • 1
  • Hongqiao Chen
    • 1
  • Qian Li
    • 1
  • Xuelian Zheng
    • 1
  • Yiping Qi
    • 2
    • 3
  • Yong Zhang
    • 1
    Email author
  1. 1.Department of Biotechnology, School of Life Science and Technology, Center for Informational BiologyUniversity of Electronic Science and Technology of ChinaChengduChina
  2. 2.Department of Plant Science and Landscape ArchitectureUniversity of MarylandCollege ParkUSA
  3. 3.Institute for Bioscience and Biotechnology ResearchUniversity of MarylandRockvilleUSA

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