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Targeting dCas9‐SunTag to a Susceptibility Gene Promoter Is Sufficient for CRISPR Interference

Title: Targeting dCas9‐SunTag to a Susceptibility Gene Promoter Is Sufficient for CRISPR Interference
Authors: Zuh‐Jyh Daniel Lin; Gabriela L. Hernandez; Myia K. Stanton; Xingguo Zheng; Kerrigan B. Gilbert; Kira M. Veley; Greg Jensen; Marisa Yoder; Suhua Feng; Basudev Ghoshal; Jason Gardiner; Ming Wang; Steven E. Jacobsen; James C. Carrington; Rebecca S. Bart
Source: Plant Direct, Vol 10, Iss 2, Pp n/a-n/a (2026)
Publisher Information: Wiley
Publication Year: 2026
Collection: Directory of Open Access Journals: DOAJ Articles
Subject Terms: biotechnology; CRISPR; gene silencing; plant disease; Botany; QK1-989
Description: Cassava production in sub‐Saharan Africa is severely impacted by diseases. Most pathogens require interaction with host susceptibility factors to complete their life cycles and cause disease. Targeted DNA methylation is an epigenetic strategy to alter gene expression in plants, and we previously reported that a zinc‐finger fused to DMS3 could establish methylation at the promoter of MeSWEET10a, a bacterial susceptibility gene, and this resulted in decreased disease. Here, we attempt a similar strategy for cassava brown streak disease. This disease is caused by the ipomoviruses CBSV and UCBSV. These viruses belong to the family Potyviridae, which has been shown extensively to require host eIF4E‐family proteins to infect plants and cause disease. We previously found that cassava plants with simultaneous knockout mutations in two eIF4E genes, nCBP‐1 and nCBP‐2, resulted in decreased susceptibility to CBSD. Here, we report successful simultaneous targeting of both promoters with methylation using a dCas9‐DRMcd‐SunTag system. However, in contrast to our previous work with MeSWEET10a, controls indicate that CRISPR interference is occurring in these lines and is sufficient for the reduction of gene expression. Future research will use genetic crosses to segregate away the DNA methylation reagents and, if DNA methylation proves heritable, assess whether methylation alone is sufficient to increase resistance to CBSD.
Document Type: article in journal/newspaper
Language: English
Relation: https://doi.org/10.1002/pld3.70128; https://doaj.org/toc/2475-4455; https://doaj.org/article/4b3fa3aaaaa34846be9483a5d2395af3
DOI: 10.1002/pld3.70128
Availability: https://doi.org/10.1002/pld3.70128; https://doaj.org/article/4b3fa3aaaaa34846be9483a5d2395af3
Accession Number: edsbas.DB108FA5
Database: BASE