A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants

  • Xu Tang
  • Zhaohui Zhong
  • Qiurong Ren
  • Binglin Liu
  • Yong ZhangEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)


The CRISPR-Cas9 system has been widely adopted in genome editing. By changing the 20 bp guide sequence, it can easily edit any sequence adjacent to a protospacer adjacent motif (PAM) in a genome. Multiplex genome editing could be accomplished with simultaneous expression of multiple single-guide RNAs (sgRNA). Given that sgRNAs are expressed by Pol III promoters, multiplex genome editing is conventionally done by assembly of multiple complete sgRNA expression cassettes together, which can be a challenge in vector construction. Here, we described a multiplex genome editing system based on a single transcript unit CRISPR-Cas9 (STU CRISPR-Cas9) expression system driven by a single Pol II promoter. It represents a novel approach for multiplex genome editing.

Key words

STU CRISPR-Cas9 Multiplex genome editing 



This work was supported by grants to YZ from the Sichuan Youth Science and Technology Foundation (2017JQ0005), the National Science Foundation of China (31771486), the National Transgenic Major Project (2018ZX08022001-003), and the Fundamental Research Funds for the Central Universities (ZYGX2016J119 and ZYGX2016J122).


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Copyright information

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

Authors and Affiliations

  • Xu Tang
    • 1
  • Zhaohui Zhong
    • 1
  • Qiurong Ren
    • 1
  • Binglin Liu
    • 1
  • 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

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