ArticleGenome research2013
Increasing frequencies of site-specific mutagenesis and gene targeting in Arabidopsis by manipulating DNA repair pathways.
Article in Genome research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 90 papers.
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Who cites it
90 citing papers in PubMed, 171 citations in OpenAlex.
- Writing Big in Plant Genomes: Advances, Challenges and Strategies for Targeted Large-Fragment DNA Insertion.Plant, cell & environment · 2026Review
- CRISPR-Based genome editing in pulses: current approaches, challenges, and future prospects.Plant molecular biology · 2025Review
- Manipulating alternative end-joining alters carbon-ion beam-induced genome mutation profiles in Arabidopsis thaliana.DNA research : an international journal for rapid publication of reports on genes and genomes · 2025Article
- CRISPR-Cas applications in agriculture and plant research.Nature reviews. Molecular cell biology · 2025Review
- Cas9-Rep fusion tethers donor DNA in vivo and boosts the efficiency of HDR-mediated genome editing.Plant biotechnology journal · 2025Article
- Review
- Beyond a few bases: methods for large DNA insertion and gene targeting in plants.The Plant journal : for cell and molecular biology · 2025Review
- Recent advances in site-specific transgene insertion into the maize genome using recombinases and genome editing endonucleases.Frontiers in plant science · 2025Review
- A long journey towards genome editing technologies in plants: a technical and critical review of genome editing technologies.Frontiers in genome editing · 2025Review
- Enhancing CRISPR-Cas-based gene targeting in tomato using a dominant-negativeHorticulture research · 2025Article
- The arsenic bioremediation using genetically engineered microbial strains on aquatic environments: An updated overview.Heliyon · 2024Review
- Identification of Site in theGenes · 2024Article
- Genetic dissection of mutagenic repair and T-DNA capture at CRISPR-induced DNA breaks inPNAS nexus · 2024Article
- Advancements in genome editing tools for genetic studies and crop improvement.Frontiers in plant science · 2024Review
- Orthologs of Human-Disease-Associated Genes in Plants Are Involved in Regulating Leaf Senescence.Life (Basel, Switzerland) · 2023Article
- Comparative analysis of seed and seedling irradiation with gamma rays and carbon ions for mutation induction in Arabidopsis.Frontiers in plant science · 2023Article
- Nonhomologous end joining as key to CRISPR/Cas-mediated plant chromosome engineering.Plant physiology · 2022Article
- Review
- Precise exogenous insertion and sequence replacements in poplar by simultaneous HDR overexpression and NHEJ suppression using CRISPR-Cas9.Horticulture research · 2022Article
- Enhancing HR Frequency for Precise Genome Editing in Plants.Frontiers in plant science · 2022Article
30 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
8 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Improved methods for engineering sequence-specific nucleases, including zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs), have made it possible to precisely modify plant genomes. However, the success of genome modification is largely dependent on the intrinsic activity of the engineered nucleases. In this study, we sought to enhance ZFN-mediated targeted mutagenesis and gene targeting (GT) in Arabidopsis by manipulating DNA repair pathways. Using a ZFN that creates a double-strand break (DSB) at the endogenous ADH1 locus, we analyzed repair outcomes in the absence of DNA repair proteins such as KU70 and LIG4 (both involved in classic nonhomologous end-joining, NHEJ) and SMC6B (involved in sister-chromatid-based homologous recombination, HR). We achieved a fivefold to 16-fold enhancement in HR-based GT in a ku70 mutant and a threefold to fourfold enhancement in GT in the lig4 mutant. Although the NHEJ mutagenesis frequency was not significantly changed in ku70 or lig4, DNA repair was shifted to microhomology-dependent alternative NHEJ. As a result, mutations in both ku70 and lig4 were predominantly large deletions, which facilitates easy screening for mutations by PCR. Interestingly, NHEJ mutagenesis and GT at the ADH1 locus were enhanced by sixfold to eightfold and threefold to fourfold, respectively, in a smc6b mutant. The increase in NHEJ-mediated mutagenesis by loss of SMC6B was further confirmed using ZFNs that target two other Arabidopsis genes, namely, TT4 and MPK8. Considering that components of DNA repair pathways are highly conserved across species, mutations in DNA repair genes likely provide a universal strategy for harnessing repair pathways to achieve desired targeted genome modifications.
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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.