CRISPR-Cas9 based gene editing is a powerful tool which can induce heritable mutations at desired genomic loci without leaving transgenic trace in plants. Here we constructed a robust, multiplex CRISPR/Cas9 system, including 12 intermediate sgRNA vectors and 6 binary vectors (https://www.addgene.org/Yao-Guang_Liu/), to accommodate diversified demanding in plants. We further developed an integrated CRISPR-GE web-based toolkit (http://skl.scau.edu.cn/) to fulfill bio-informatic processing and analysis requirements while using CRISPR. This CRISPR vector system has been applied in many crops including rice. As an example, we created glutinous rice with the transgene cassette segregated within two generations using our vector. This high efficiency, and user-friendly CRISPR platform are benefiting plant research and crop improvements.
1. Introducing the multiplex CRISPR-Cas9 vector system
2. CRISPR-GE web based toolkit for designing CRISPR experiment and analyzing mutants in plant