ArticleNucleic acids research2019
Targeted DNA transposition in vitro using a dCas9-transposase fusion protein.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
20 citing papers in PubMed.
- Structure-based recasting of a mammalian DNA transpososome as an obligate heterodimer.bioRxiv : the preprint server for biology · 2026Article
- Precision gene editing: The power of CRISPR-Cas in modern genetics.Molecular therapy. Nucleic acids · 2025Review
- Exploration of the potential of genomic editing in the treatment of congenital adrenal hyperplasia.Frontiers in endocrinology · 2025Review
- From bench to bedside: cutting-edge applications of base editing and prime editing in precision medicine.Journal of translational medicine · 2024Review
- Next-generation CRISPR technology for genome, epigenome and mitochondrial editing.Transgenic research · 2024Review
- Natural and Engineered Guide RNA-Directed Transposition with CRISPR-Associated Tn7-Like Transposons.Annual review of biochemistry · 2024Review
- Article
- Applications and Research Advances in the Delivery of CRISPR/Cas9 Systems for the Treatment of Inherited Diseases.International journal of molecular sciences · 2023Review
- Recent advances in therapeutic CRISPR-Cas9 genome editing: mechanisms and applications.Molecular biomedicine · 2023Review
- Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases.Nature biotechnology · 2023Article
- Strategies to improve homology-based repair outcomes following CRISPR-based gene editing in mosquitoes: lessons in how to keep any repair disruptions local.Virology journal · 2022Review
- Genetically engineered bacterium: Principles, practices, and prospects.Frontiers in microbiology · 2022Review
- Integration of diverse DNA substrates by a casposase can be targeted to R-loops in vitro by its fusion to Cas9.Bioscience reports · 2021Article
- Insertion Specificity of the hATx-6 Transposase ofFrontiers in molecular biosciences · 2021Article
- Enhancing site-specific DNA integration by a Cas9 nuclease fused with a DNA donor-binding domain.Nucleic acids research · 2020Article
- Potential of transposon-mediated cellular reprogramming towards cell-based therapies.World journal of stem cells · 2020Review
- RNA-guided retargeting of SeLife · 2020Article
- Jump around: transposons in and out of the laboratory.F1000Research · 2020Review
- An Engineered Cas-Transposon System for Programmable and Site-Directed DNA Transpositions.The CRISPR journal · 2019Article
- A highly soluble Sleeping Beauty transposase improves control of gene insertion.Nature biotechnology · 2019Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
2 authors.
Funding
Abstract
Homology-directed genome engineering is limited by transgene size. Although DNA transposons are more efficient with large transgenes, random integrations are potentially mutagenic. Here we present an in vitro mechanistic study that demonstrates efficient Cas9 targeting of the mariner transposon Hsmar1. Integrations were unidirectional and tightly constrained to one side of the sgRNA binding site. Further analysis of the nucleoprotein intermediates demonstrated that the transposase and Cas9 moieties can bind their respective substrates independently or in concert. Kinetic analysis of the reaction in the presence of the Cas9 target-DNA revealed a delay between first and second strand cleavage at the transposon end. This step involves a significant conformational change that may be hindered by the properties of the interdomainal linker. Otherwise, the transposase moiety behaved normally and was proficient for integration in vitro and in Escherichia coli. Specific integration into the lacZ gene in E. coli was obscured by a high background of random integrations. Nevertheless, Cas9 is an attractive candidate for transposon-targeting because it has a high affinity and long dwell-time at its target site. This will facilitate a future optogenetic strategy for the temporal control of integration, which will increase the ratio of targeted to untargeted events.
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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.