ArticleScientific reports2018
Highly efficient heritable targeted deletions of gene clusters and non-coding regulatory regions in Arabidopsis using CRISPR/Cas9.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 86 citations in OpenAlex.
- iNOME-seq: in vivo simultaneous genome-wide mapping of chromatin accessibility, nucleosome positioning, DNA-binding protein sites, and DNA methylation in Arabidopsis.Genome biology · 2025Article
- Targeted deletions of large syntenic regions inProceedings of the National Academy of Sciences of the United States of America · 2025Article
- A CRISPR/Cas9-based vector system enables the fast breeding of selection-marker-free canola with Rcr1-rendered clubroot resistance.Journal of experimental botany · 2024Article
- Long noncoding RNA-mediated epigenetic regulation of auxin-related genes controls shade avoidance syndrome in Arabidopsis.The EMBO journal · 2023Article
- Hidden prevalence of deletion-inversion bi-alleles in CRISPR-mediated deletions of tandemly arrayed genes in plants.Nature communications · 2023Article
- An actin remodeling role for Arabidopsis processing bodies revealed by their proximity interactome.The EMBO journal · 2023Article
- Targeted gene deletion with SpCas9 and multiple guide RNAs in Arabidopsis thaliana: four are better than two.Plant methods · 2023Article
- Engineering the Biosynthesis of Late-Stage Vinblastine Precursors Precondylocarpine Acetate, Catharanthine, Tabersonine inACS synthetic biology · 2023Article
- A native promoter-gene fusion created by CRISPR/Cas9-mediated genomic deletion offers a transgene-free method to drive oil accumulation in leaves.FEBS letters · 2022Article
- CRISPR-Cas9-mediated chromosome engineering in Arabidopsis thaliana.Nature protocols · 2022Review
- Novel Plant Breeding Techniques Shake Hands with Cereals to Increase Production.Plants (Basel, Switzerland) · 2022Review
- Nonhomologous end joining as key to CRISPR/Cas-mediated plant chromosome engineering.Plant physiology · 2022Article
- Photosynthetic Light Harvesting and Thylakoid Organization in a CRISPR/Cas9 Arabidopsis Thaliana LHCB1 Knockout Mutant.Frontiers in plant science · 2022Article
- RNA Pol III promoters-key players in precisely targeted plant genome editing.Frontiers in genetics · 2022Review
- Genome editing with CRISPR/Cas9 in Pinus radiata (D. Don).BMC plant biology · 2021Article
- Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Multiplex Genome-Editing Technologies for Revolutionizing Plant Biology and Crop Improvement.Frontiers in plant science · 2021Review
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- Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of HeritableBio-protocol · 2020Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
Abstract
Genome editing using CRISPR/Cas9 is considered the best instrument for genome engineering in plants. This methodology is based on the nuclease activity of Cas9 that is guided to specific genome sequences by single guide RNAs (sgRNAs) thus enabling researchers to engineer simple mutations or large chromosomal deletions. Current methodologies for targeted genome editing in plants using CRISPR/Cas9 are however largely inefficient, mostly due to low Cas9 activity, variable sgRNA efficiency and low heritability of genetic lesions. Here, we describe a newly developed strategy to enhance CRISPR/Cas9 efficiency in Arabidopsis thaliana focusing on the design of novel binary vectors (pUbiCAS9-Red and pEciCAS9-Red), the selection of highly efficient sgRNAs, and the use of direct plant regeneration from induced cell cultures. Our work demonstrates that by combining these three independent developments, heritable targeted chromosomal deletions of large gene clusters and intergenic regulatory sequences can be engineered at a high efficiency. Our results demonstrate that this improved CRISPR/Cas9 methodology can provide a fast, efficient and cost-effective tool to engineer targeted heritable chromosomal deletions, which will be instrumental for future high-throughput functional genomics studies in plants.
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