ArticleCommunications biology2022
Efficient modification and preparation of circular DNA for expression in cell culture.
Article in Communications biology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 17 citations in OpenAlex.
- Overestimations in the cyclization efficiency of small, restriction enzyme-digested DNA fragments.Nature protocols · 2026Article
- Reply to: Overestimations in the cyclization efficiency of small, restriction enzyme-digested DNA fragments.Nature protocols · 2026Article
- DNA Cut-Ligation Cyclization Surpasses Jacobson-Stockmayer J-Factor Expectations by over Threefold.Biomolecules · 2026Article
- CD3ɛ Nanobody-Engineered Extracellular Vesicles Driving In Vivo Generation of TCE-secreting CAR-Ts for Solid Tumor Therapy With Memory Response and Minimal Immunogenicity.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Plasmid2MC: efficient cell-free generation of high-purity minicircle DNA for genome editing in mammalian cells.Communications biology · 2025Article
- TOPO-seq reveals DNA topology-induced off-target activity by Cas9 and base editors.Nature chemical biology · 2025Article
- Circular Vectors as an efficient, fully synthetic, cell-free approach for preparing small circular DNA as a plasmid substitute for guide RNA expression in CRISPR-Cas9 genome editing.Nature protocols · 2025Review
- Synthesizing supercoiled circular DNA molecules in vitro.Nucleic acids research · 2025Article
- How many plasmids can bacteria carry? A synthetic biology perspective.Open biology · 2025Review
- Tissue nanotransfection and cellular reprogramming in regenerative medicine and antimicrobial dynamics.Frontiers in bioengineering and biotechnology · 2025Review
- Extrachromosomal circular DNA drives dynamic genome plasticity: emerging roles in disease progression and clinical potential.Theranostics · 2025Review
- Continuous evolution of user-defined genes at 1 million times the genomic mutation rate.Science (New York, N.Y.) · 2024Article
- Established and Emerging Methods for Protecting Linear DNA in Cell-Free Expression Systems.Methods and protocols · 2023Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
DNA plasmids are an essential tool for delivery and expression of RNAs and proteins in cell culture experiments. The preparation of plasmids typically involves a laborious process of bacterial cloning, validation, and purification. While the expression plasmids can be designed and ordered from the contract manufacturers, the cost may be prohibitive when a large number of plasmids is required. We have developed an efficient fully synthetic method and protocol that enables the production of circularized DNA containing expression elements ready for transfection in as little as 3 hours, thereby eliminating the bacterial cloning steps. The protocol describes how to take a linear double-stranded DNA fragment and efficiently circularize and purify this DNA fragment with minimal hands-on time. As proof of the principle, we applied Circular Vector expressing engineered prime editing guide RNA (epegRNA) in cell culture, and demonstrated matching and even exceeding performance of this method as compared to guides expressed by plasmids. The method's speed of preparation, low cost, and ease of use will make it a useful tool in applications requiring the expression of short RNAs and proteins.
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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.