ArticlePloS one2019
Multiplexed editing of a begomovirus genome restricts escape mutant formation and disease development.
Article in PloS one, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 75 citations in OpenAlex.
- Leaf Curl Disease Resistance Landscaping in Homozygous Ty-Gene Donor Tomato Genotypes Using a Robust Disease Scoring System and Indexing of Begomoviruses Under Natural Epiphytotic Conditions.Current microbiology · 2026Article
- Enhanced Rice Yellow Mottle Virus Resistance via CRISPR/Cas9-Targeted Mutagenesis of the Rice eIF(iso)4G Gene.Molecular plant pathology · 2026Article
- Evaluation of CRISPR/Cas9 based single and multiplex gRNAs against ChiLCV in tobacco.Journal, genetic engineering & biotechnology · 2026Article
- Advances in CRISPR Plant Applications.International journal of molecular sciences · 2026Review
- Genome editing‑based strategies to combat geminiviruses: CRISPR/Cas9 and emerging high‑fidelity tools.Archives of microbiology · 2026Review
- Geminiviral-CR-gRNA expressed in cowpea efficiently edited MYMV and MYMIV genome to provide resistance against cowpea yellow mosaic disease without hampering plant growth and yield.BMC plant biology · 2026Article
- Strategies for plant-virus disease management from gene editing to nanotechnology.Physiology and molecular biology of plants : an international journal of functional plant biology · 2025Review
- Potential for Duplexed, In-Tandem gRNA-Mediated Suppression of Two Essential Genes of Tomato Leaf Curl New Delhi Virus in Crop Plants.Pathogens (Basel, Switzerland) · 2025Article
- Molecular and genomic insights into viral resistance inFrontiers in plant science · 2025Review
- RPA-assisted CRISPR-Cas12a-enabled point-of-care diagnostic platform for chili leaf curl virus with fluorescent and colorimetric readouts.Frontiers in microbiology · 2025Article
- CRISPR/Cas: An Emerging Toolbox for Engineering Virus Resistance in Plants.Plants (Basel, Switzerland) · 2024Review
- Article
- Emerging evidence of seed transmission of begomoviruses: implications in global circulation and disease outbreak.Frontiers in plant science · 2024Review
- CRISPR/Cas9-gene editing approaches in plant breeding.GM crops & food · 2023Review
- Targeted mutagenesis with sequence-specific nucleases for accelerated improvement of polyploid crops: Progress, challenges, and prospects.The plant genome · 2023Review
- Development of Highly Efficient Resistance toInternational journal of molecular sciences · 2023Article
- Current status, breeding strategies and future prospects for managing chilli leaf curl virus disease and associated begomoviruses in Chilli (Frontiers in plant science · 2023Review
- Genome editing for healthy crops: traits, tools and impacts.Frontiers in plant science · 2023Review
- Genome editing (CRISPR-Cas)-mediated virus resistance in potato (Solanum tuberosum L.).Molecular biology reports · 2022Review
- CRISPR/Cas-based tools for the targeted control of plant viruses.Molecular plant pathology · 2022Review
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Whitefly-transmitted begomoviruses cause serious damage to many economically important food, feed, and fiber crops. Numerous vegetable crops are severely affected and chilli leaf curl virus (ChiLCV) is the most dominant and widely distributed begomovirus in chilli (Capsicum annuum) throughout the Indian subcontinent. Recently, CRISPR-Cas9 technology was used as a means to reduce geminivirus replication in infected plants. However, this approach was shown to have certain limitations such as the evolution of escape mutants. In this study, we used a novel, multiplexed guide RNA (gRNA) based CRISPR-Cas9 approach that targets the viral genome at two or more sites simultaneously. This tactic was effective in eliminating the ChiLCV genome without recurrence of functional escape mutants. Six individual gRNA spacer sequences were designed from the ChiLCV genome and in vitro assays confirmed the cleavage behaviour of these spacer sequences. Multiplexed gRNA expression clones, based on combinations of the above-mentioned spacer sequences, were developed. A total of nine-duplex and two-triplex CRISPR-Cas9 constructs were made. The efficacy of these constructs was tested for inhibition of ChiLCV infection in Nicotiana benthamiana. Results indicated that all the constructs caused a significant reduction in viral DNA accumulation. In particular, three constructs (gRNA5+4, gRNA5+2 and gRNA1+2) were most effective in reducing the viral titer and symptoms. T7E1 assay and sequencing of the targeted viral genome did not detect any escape mutants. The multiplexed genome-editing technique could be an effective way to trigger a high level of resistance against begemoviruses. To our knowledge, this is the first report of demonstrating the effectiveness of a multiplexed gRNA-based plant virus genome editing to minimize and eliminate escape mutant formation.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.