ArticlePlant physiology2016
Biallelic Gene Targeting in Rice.
Article in Plant physiology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 81 papers.
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Who cites it
81 citing papers in PubMed, 202 citations in OpenAlex.
- SSA-mediated selection marker gene activation enhances relative gene targeting efficiency in plants.Horticulture research · 2025Article
- Generating Broad-Spectrum Resistance to ALS-Inhibiting Herbicides in Rice by CRISPR/Cas9-Mediated NHEJ.Rice (New York, N.Y.) · 2025Article
- CRISPR/Cas-mediated genome editing: playing a versatile role in mitigating the challenges of sustainable rice improvement.3 Biotech · 2025Review
- Enhancing Specificity, Precision, Accessibility, Flexibility, and Safety to Overcome Traditional CRISPR/Cas Editing Challenges and Shape Future Innovations.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Review
- Cas9-Rep fusion tethers donor DNA in vivo and boosts the efficiency of HDR-mediated genome editing.Plant biotechnology journal · 2025Article
- Beyond a few bases: methods for large DNA insertion and gene targeting in plants.The Plant journal : for cell and molecular biology · 2025Review
- Development of efficient targeted insertion mediated by CRISPR-Cas12a and homology-directed repair in maize.Frontiers in genome editing · 2025Article
- Enhancing CRISPR-Cas-based gene targeting in tomato using a dominant-negativeHorticulture research · 2025Article
- The Potential of CRISPR/Cas9 to Circumvent the Risk Factor NeurotoxinCurrent issues in molecular biology · 2024Review
- Review
- Precise genetic engineering withPlant biotechnology (Tokyo, Japan) · 2023Article
- Excision of DNA fragments with thePlant biotechnology (Tokyo, Japan) · 2023Article
- Knockout Mutants ofInternational journal of molecular sciences · 2023Article
- Genetic basis controlling rice plant architecture and its modification for breeding.Breeding science · 2023Article
- In Vivo Genome Editing in Type I and II Methanotrophs Using a CRISPR/Cas9 System.ACS synthetic biology · 2023Article
- Precise and heritable gene targeting in rice using a sequential transformation strategy.Cell reports methods · 2023Article
- Green revolution to genome revolution: driving better resilient crops against environmental instability.Frontiers in genetics · 2023Review
- Multi-allelic gene editing in an apomictic, tetraploid turf and forage grass (Frontiers in plant science · 2023Article
- Methods of crop improvement and applications towards fortifying food security.Frontiers in genome editing · 2023Review
- CRISPR-Cas9-based precise engineering of SlHyPRP1 protein towards multi-stress tolerance in tomato.Frontiers in plant science · 2023Article
21 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Sequence-specific nucleases (SSNs) have been used successfully in homology-directed repair (HDR)-mediated gene targeting (GT) in many organisms. However, break-induced GT in plants remains challenging due to inefficient delivery of HDR templates and SSNs into plant nuclei. In many plants, including rice, Agrobacterium-mediated transformation is the most practical means of transformation because this biotic transformation system can deliver longer and more intact DNA payloads with less incorporation of fragmented DNA compared with physical transformation systems such as polyethylene glycol, electroporation, or biolistics. Following infection with Agrobacterium, transfer of transfer DNA (T-DNA) to the nucleus and its integration into the plant genome occur consecutively during cocultivation, thus timing the induction of DNA double-strand breaks (DSBs) on the target gene to coincide with the delivery of the HDR template is crucial. To synchronize DSB induction and delivery of the HDR template, we transformed a Cas9 expression construct and GT vector harboring the HDR template with guide RNAs (gRNAs) targeting the rice acetolactate synthase (ALS) gene either separately or sequentially into rice calli. When gRNAs targeting ALS were transcribed transiently from double-stranded T-DNA containing the HDR template, DSBs were induced in the ALS locus by the assembled Cas9/gRNA complex and homologous recombination was stimulated. Contrary to our expectations, there was no great difference in GT efficiency between Cas9-expressing and nonexpressing cells. However, when gRNA targeting DNA ligase 4 was transformed with Cas9 prior to the GT experiment, GT efficiency increased dramatically and more than one line exhibiting biallelic GT at the ALS locus was obtained.
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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.