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  • Title: A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants.
    Author: Tang X, Zhong Z, Ren Q, Liu B, Zhang Y.
    Journal: Methods Mol Biol; 2019; 1917():75-82. PubMed ID: 30610629.
    Abstract:
    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.
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