ArticlePlant molecular biology2020
Development of multiplex genome editing toolkits for citrus with high efficacy in biallelic and homozygous mutations.
Article in Plant molecular biology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 72 citations in OpenAlex.
- REF1 peptides significantly enhance transformation efficiency but inhibit shoot regeneration in citrusHorticulture research · 2026Article
- Induction of Embryogenic Callus, Protoplast Isolation, and PEG-Mediated Transformation Protocols inPlants (Basel, Switzerland) · 2026Article
- Cas9/sgRNA-mediated genome editing of citrus via mature tissue transformation enables both high-efficacy genome editing and early flowering.Frontiers in plant science · 2026Article
- The Enhancement of Fungal Disease Resistance in Major Staple Crops Using CRISPR-Cas Technology.Genes · 2025Review
- Genetic regulations of citrus flowering: insights towards climatic factors and modern biotechnological approaches.Planta · 2025Review
- Substantial enhancement ofHorticulture research · 2025Article
- In planta genome editing in citrus facilitated by co-expression of CRISPR/Cas and developmental regulators.The Plant journal : for cell and molecular biology · 2025Article
- Construction and Validation of CRISPR/Cas Vectors for Editing theCurrent issues in molecular biology · 2024Article
- An efficient multiplex approach to CRISPR/Cas9 gene editing in citrus.Plant methods · 2024Article
- Review
- Cas12a RNP-mediated co-transformation enables transgene-free multiplex genome editing, long deletions, and inversions in citrus chromosome.Frontiers in plant science · 2024Article
- Generation of transgene-free canker-resistantFrontiers in plant science · 2024Article
- Genome editing in East African cichlids and tilapias: state-of-the-art and future directions.Open biology · 2023Review
- Transgene-free genome editing of vegetatively propagated and perennial plant species in the T0 generation via a co-editing strategy.Nature plants · 2023Article
- Generation of the transgene-free canker-resistant Citrus sinensis using Cas12a/crRNA ribonucleoprotein in the T0 generation.Nature communications · 2023Article
- Knockout of floral and meiosis genes using CRISPR/Cas9 produces male-sterility in Eucalyptus without impacts on vegetative growth.Plant direct · 2023Article
- Citrus genomic resources unravel putative genetic determinants of Huanglongbing pathogenicity.iScience · 2023Article
- Applications of CRISPR/Cas genome editing in economically important fruit crops: recent advances and future directions.Molecular horticulture · 2023Review
- CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane.Frontiers in plant science · 2023Review
- Molecular Aspects of MicroRNAs and Phytohormonal Signaling in Response to Drought Stress: A Review.Current issues in molecular biology · 2022Review
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
key messageWe have developed multiplex genome editing toolkits for citrus that significantly improve citrus genome editing efficacy. CRISPR/Cas systems have been engineered for genome editing in many organisms, including plants. However, the gene editing efficiency in citrus via CRISPR technology remains too low to be implemented for genetic improvement in practice. Moreover, it is very difficult to obtain homozygous or biallelic knockout mutants in citrus. Here, we have developed multiplex genome editing toolkits for citrus including PEG-mediated protoplast transformation, a GFP reporter system that allows the rapid assessment of CRISPR constructs, citrus U6 promoters with improved efficacy, and tRNA-mediated or Csy4-mediated multiplex genome editing. Using the toolkits, we successfully conducted genome modification of embryogenic protoplast cells and epicotyl tissues. We have achieved a biallelic mutation rate of 44.4% and a homozygous mutation rate of 11.1%, representing a significant improvement in citrus genome editing efficacy. In addition, our study lays the foundation for nontransgenic genome editing of citrus.
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Registered trials
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.