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

Yingxiao Zhang; Yong Zhang; Yiping Qi

doi:10.1007/978-1-4939-8991-1_18

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

Methods Mol Biol. 2019:1917:245-256. doi: 10.1007/978-1-4939-8991-1_18.

Authors

Yingxiao Zhang¹, Yong Zhang², Yiping Qi^{3

4}

Affiliations

¹ Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA.
² Department of Biotechnology, School of Life Science and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Chengdu, China.
³ Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD, USA. yiping@umd.edu.
⁴ Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD, USA. yiping@umd.edu.

PMID: 30610641
DOI: 10.1007/978-1-4939-8991-1_18

Abstract

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.

Keywords: CRISPR-Cpf1 (Cas12a); Gateway cloning; Plant gene knockdown; Plant gene knockout.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / genetics
CRISPR-Cas Systems / genetics*
Gene Editing
Gene Knockout Techniques / methods*
Oryza / genetics