Using CRISPR/Cas9 system to introduce targeted mutation in Arabidopsis

Abstract

Gene editing enables the modification of genetic information in a targeted manner. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) is a gene editing approach that has been widely used in both the animal and plant studies. Plant transcription factors play critical roles in plant development. Recent studies have used the CRISPER/Cas9 system to disrupt transcription factors and cis-elements in plants. In this chapter, we describe a protocol of using the CRISPR/Cas9 system to generate transcription factor mutants in the model plant species Arabidopsis thaliana. Our protocol contains five different steps: (1) adaptor primers design, (2) cloning of sgRNA expression cassettes, (3) Golden Gate cloning, (4) E. coli and Agrobacterium transformation, and (5) Arabidopsis transformation.