A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants
The CRISPR-Cas9 system has been widely adopted in genome editing. By changing the 20 bp guide sequence, it can easily edit any sequence adjacent to a protospacer adjacent motif (PAM) in a genome. Multiplex genome editing could be accomplished with simultaneous expression of multiple single-guide RNAs (sgRNA). Given that sgRNAs are expressed by Pol III promoters, multiplex genome editing is conventionally done by assembly of multiple complete sgRNA expression cassettes together, which can be a challenge in vector construction. Here, we described a multiplex genome editing system based on a single transcript unit CRISPR-Cas9 (STU CRISPR-Cas9) expression system driven by a single Pol II promoter. It represents a novel approach for multiplex genome editing.
Key wordsSTU CRISPR-Cas9 Multiplex genome editing
This work was supported by grants to YZ from the Sichuan Youth Science and Technology Foundation (2017JQ0005), the National Science Foundation of China (31771486), the National Transgenic Major Project (2018ZX08022001-003), and the Fundamental Research Funds for the Central Universities (ZYGX2016J119 and ZYGX2016J122).
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