4.3 Article

Gene tagging via CRISPR-mediated homology-directed repair in cassava

Journal

G3-GENES GENOMES GENETICS
Volume 11, Issue 4, Pages -

Publisher

OXFORD UNIV PRESS INC
DOI: 10.1093/g3journal/jkab028

Keywords

bacterial pathogens; pathogenesis; genome editing; Xanthomonas; cassava

Funding

  1. Bill and Melinda Gates Foundation [OPP1125410]
  2. National Science Foundation (NSF EDGE Award) [1827761]

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Research has focused on visualizing the initial steps of Cassava Bacterial Blight infection and successfully demonstrated HDR in cassava through gene editing. This could help address the bottleneck in crop improvement.
Research on a few model plant-pathogen systems has benefitted from years of tool and resource development. This is not the case for the vast majority of economically and nutritionally important plants, creating a crop improvement bottleneck. Cassava bacterial blight (CBB), caused by Xanthomonas axonopodis pv. manihotis (Xam), is an important disease in all regions where cassava (Manihot esculenta Crantz) is grown. Here, we describe the development of cassava that can be used to visualize one of the initial steps of CBB infection in vivo. Using CRISPR-mediated homology-directed repair (HDR), we generated plants containing scarless insertion of GFP at the 3' end of CBB susceptibility (S) gene MeSWEET10a. Activation of MeSWEET10a-GFP by the transcription activator-like (TAL) effector TAL20 was subsequently visualized at transcriptional and translational levels. To our knowledge, this is the first such demonstration of HDR via gene editing in cassava.

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