ArticleNucleic acids research2003
DNA modification and functional delivery into human cells using Escherichia coli DH10B.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 60 citations in OpenAlex.
- Engineered bacteria launch and control an oncolytic virus.Nature biomedical engineering · 2026Article
- Synthetic rewriting technologies in mammalian cells.Nature communications · 2026Review
- Comparison of the virulence of community- and hospital- isolated Acinetobacter baumannii in HeLa cell line and insect model, Galleria mellonella.Access microbiology · 2025Article
- Evolution and synthetic biology.Current opinion in microbiology · 2023Review
- A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds.Micromachines · 2017Article
- Genetic Stability of Bacterial Artificial Chromosome-Derived Human Cytomegalovirus during Culture In Vitro.Journal of virology · 2016Article
- A pathway from chromosome transfer to engineering resulting in human and mouse artificial chromosomes for a variety of applications to bio-medical challenges.Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology · 2015Review
- Article
- Human artificial chromosomes for gene delivery and the development of animal models.Molecular therapy : the journal of the American Society of Gene Therapy · 2011Review
- Proliferation behavior of E. coli in a three-dimensional in vitro tumor model.Integrative biology : quantitative biosciences from nano to macro · 2011Article
- Site-specific chromosomal integration of large synthetic constructs.Nucleic acids research · 2010Article
- Prospects for the use of artificial chromosomes and minichromosome-like episomes in gene therapy.Journal of biomedicine & biotechnology · 2010Article
- Recombination between linear double-stranded DNA substrates in vivo.Analytical biochemistry · 2009Article
- Bacterial conjugation in the cytoplasm of mouse cells.Infection and immunity · 2008Article
- An intronic sequence mutated in flexed-tail mice regulates splicing of Smad5.Mammalian genome : official journal of the International Mammalian Genome Society · 2007Article
- Bacterial transfer of large functional genomic DNA into human cells.Gene therapy · 2005Article
- Artificial and engineered chromosomes: developments and prospects for gene therapy.Chromosoma · 2005Review
- A place for everything: chromosomal integration of large constructs.Bioengineered bugsArticle
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
The availability of almost the complete human genome as cloned BAC libraries represents a valuable resource for functional genomic analysis, which, however, has been somewhat limited by the ability to modify and transfer this DNA into mammalian cells intact. Here we report a novel comprehensive Escherichia coli-based vector system for the modification, propagation and delivery of large human genomic BAC clones into mammalian cells. The GET recombination inducible homologous recombination system was used in the BAC host strain E.coli DH10B to precisely insert an EGFPneo cassette into the vector portion of a approximately 200 kb human BAC clone, providing a relatively simple method to directly convert available BAC clones into suitable vectors for mammalian cells. GET recombination was also used for the targeted deletion of the asd gene from the E.coli chromosome, resulting in defective cell wall synthesis and diaminopimelic acid auxotrophy. Transfer of the Yersinia pseudotuberculosis invasin gene into E.coli DH10B asd(-) rendered it competent to invade HeLa cells and deliver DNA, as judged by transient expression of green fluorescent protein and stable neomycin-resistant colonies. The efficiency of DNA transfer and survival of HeLa cells has been optimized for incubation time and multiplicity of infection of invasive E.coli with HeLa cells. This combination of E.coli-based homologous recombination and invasion technologies using BAC host strain E.coli DH10B will greatly improve the utility of the available BAC libraries from the human and other genomes for gene expression and functional genomic studies.
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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.