Chapter title |
Using CRISPR/Cas9 System to Introduce Targeted Mutation in Arabidopsis
|
---|---|
Chapter number | 6 |
Book title |
Plant Transcription Factors
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8657-6_6 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8656-9, 978-1-4939-8657-6
|
Authors |
Ze Hong Lee, Nobutoshi Yamaguchi, Toshiro Ito, Lee, Ze Hong, Yamaguchi, Nobutoshi, Ito, Toshiro |
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. |
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