Plant Gene Knockout and Knockdown by CRISPR-Cpf1 (Cas12a) Systems

  • Yingxiao Zhang
  • Yong Zhang
  • Yiping Qi
Part of the Methods in Molecular Biology book series (MIMB, volume 1917)


CRISPR-Cpf1 (Cas12a) is a class II type V endonuclease, which has been used as a genome editing tool in different biological systems. Here we describe a fast, efficient, and user-friendly system for CRISPR-Cpf1 expression vector assembly. In this system, the Pol II promoter is used to drive the expression of both Cpf1 and its crRNA, with the crRNA flanked by hammerhead (HH) and hepatitis delta virus (HDV) ribozyme RNAs for precise crRNA processing. All the components of this system can be modified depending on plant species and experimental goals. Using this system, nearly 100% editing efficiency and 90% gene expression decrease were achieved in rice and Arabidopsis, respectively.

Key words

CRISPR-Cpf1 (Cas12a) Plant gene knockout Plant gene knockdown Gateway cloning 



This work is supported by funds from University of Maryland and Syngenta Biotechnology.


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Copyright information

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

Authors and Affiliations

  1. 1.Department of Plant Science and Landscape ArchitectureUniversity of MarylandCollege ParkUSA
  2. 2.Department of Biotechnology, School of Life Science and Technology, Center for Informational BiologyUniversity of Electronic Science and Technology of ChinaChengduChina
  3. 3.Institute for Bioscience and Biotechnology ResearchUniversity of MarylandRockvilleUSA

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