CRISPR in Plants
摘要
In this chapter, we describe several strategies for implementing CRISPR-Cas9 genome editing in Arabidopsis thaliana, a model plant. Most experimental approaches described here will take several weeks of laboratory time (assuming classes meet once per week). For a molecular biology classroom, we describe a simple method for cloning a gene knock-out guide sequence into a plant binary vector plasmid. For genetics or plant biology courses, we describe an approach starting with a plasmid vector containing a guide sequence and Cas9 and have the students engineer transgenic A. thaliana plants. A third strategy would be to have students analyze transgenic seed lines and determine the sequence of the edited region. All these approaches introduce students to TAIR (The Arabidopsis Information Resource), an invaluable resource allowing students to examine gene sequences and expression profiles for suitable genes for knock-out experiments. We also address the challenges and benefits of in planta studies.