Schematic representation of the guide selection procedure and workflow followed to evaluate the CRISPR-Cas CHLI reporter system. (IMAGE)

Nanjing Agricultural University The Academy of Science

Schematic representation of the guide selection procedure and workflow followed to evaluate the CRISPR-Cas CHLI reporter system.

Caption

Schematic representation of the guide selection procedure and workflow followed to evaluate the CRISPR-Cas CHLI reporter system. (a) Metabolic pathway of chlorophyll biosynthesis, in which CHLI is involved. Three different crRNAs were selected that can simultaneously target both homeologs of NbCHLI, named here CHLI-A and CHLI-B. Target selection was based on the specificity scores given by CRISPOR tool. (b) The crRNA was formed by the LbCas12a direct repeat (DR1) scaffold, and the 23-nt protospacer (PS). The three selected crRNAs were cloned into a TRV vector, and their expression was driven by the PEBV promoter. (c) N. benthamiana plants were agroinoculated with the three constructs, and gene editing was analyzed at 21 dpi in systemic non-inoculated leaves displaying bleaching phenotype.

Credit

Horticulture Research

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