ArticleBMC biology2024
Application of multiple sgRNAs boosts efficiency of CRISPR/Cas9-mediated gene targeting in Arabidopsis.
Article in BMC biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 29 citations in OpenAlex.
- Establishment of CRISPR/Cas9 Genome Editing inJournal of fungi (Basel, Switzerland) · 2026Article
- VEGFA-Targeted M3-F4 Ionizable Lipid Nanoparticles Improve Diabetic Retinopathy.Molecular pharmaceutics · 2026Article
- Efficient genetic perturbation of murine sensory neurons in vivo using CRISPR/Cas9.The journal of pain · 2026Article
- Dual-single-guide RNA strategy improves CRISPR-mediated homology-directed repair in Aspergillus.Nucleic acids research · 2026Article
- Synthetic Biology Strategies for Activating Cryptic BGCs inACS omega · 2026Review
- In vivo genome editing with a novel Cj4Cas9.Communications biology · 2025Article
- Technological advances in visualizing and rewiring microtubules during plant development.Journal of experimental botany · 2025Review
- SSA-mediated selection marker gene activation enhances relative gene targeting efficiency in plants.Horticulture research · 2025Article
- Efficient genetic perturbation of murine sensory neuronsbioRxiv : the preprint server for biology · 2025Article
- RNA Polymerase III Promoters Compatible with CRISPR Gene Regulation inACS synthetic biology · 2025Article
- Applying conventional and cell-type-specific CRISPR/Cas9 genome editing in legume plants.aBIOTECH · 2025Review
- Harnessing promoter elements to enhance gene editing in plants: perspectives and advances.Plant biotechnology journal · 2025Review
- An Optimized Editing Approach for Wheat Genes by Improving sgRNA Design and Transformation Strategies.International journal of molecular sciences · 2025Article
- Single-stranded DNA (ssDNA) donor repair templates and CRISPR/Cas9 enable a high-frequency of targeted insertions in potato.Frontiers in genome editing · 2025Article
- Gene Targeting via CRISPR/Cas9-Mediated DNA Double-Strand Break Induction in Rice.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Double step screening using endogenous marker improves relative gene targeting efficiency in Arabidopsis.Scientific reports · 2024Article
- Cas12a-mediated gene targeting by sequential transformation strategy in Arabidopsis thaliana.BMC plant biology · 2024Article
- Simple promotion of Cas9 and Cas12a expression improves gene targeting via an all-in-one strategy.Frontiers in plant science · 2024Article
- Advancements, challenges, and future perspectives in developing feline herpesvirus 1 as a vaccine vector.Frontiers in immunology · 2024Review
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
backgroundPrecise gene targeting (GT) is a powerful tool for heritable precision genome engineering, enabling knock-in or replacement of the endogenous sequence via homologous recombination. We recently established a CRISPR/Cas9-mediated approach for heritable GT in Arabidopsis thaliana (Arabidopsis) and rice and reported that the double-strand breaks (DSBs) frequency of Cas9 influences the GT efficiency. However, the relationship between DSBs and GT at the same locus was not examined. Furthermore, it has never been investigated whether an increase in the number of copies of sgRNAs or the use of multiple sgRNAs would improve the efficiency of GT.
resultsHere, we achieved precise GT at endogenous loci Embryo Defective 2410 (EMB2410) and Repressor of Silencing 1 (ROS1) using the sequential transformation strategy and the combination of sgRNAs. We show that increasing of sgRNAs copy number elevates both DSBs and GT efficiency. On the other hand, application of multiple sgRNAs does not always enhance GT efficiency. Our results also suggested that some inefficient sgRNAs would play a role as a helper to facilitate other sgRNAs DSBs activity.
conclusionsThe results of this study clearly show that DSB efficiency, rather than mutation pattern, is one of the most important key factors determining GT efficiency. This study provides new insights into the relationship between sgRNAs, DSBs, and GTs and the molecular mechanisms of CRISPR/Cas9-mediated GTs in plants.
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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.