Recombinant Escherichia coli as a gene delivery vector into airway epithelial cells

Fajac, Isabelle; Grosse, Stéphanie; Collombet, Jean-Marc; Thévenot, Guiti; Goussard, Sylvie; Danel, Claire; Grillot‐Courvalin, Catherine

doi:10.1016/j.jconrel.2004.03.025

Cited by 29 publications

(33 citation statements)

References 18 publications

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“…Proof-of-principle for bactofection in a range of non-phagocytic cell lines, including airway epithelial cells, has been established in vitro. 13,14,[17][18][19] These studies have mainly focused on transfer of the GFP reporter gene. However, more relevant to our work, recombinant E. coli and L. monocytogenes were recently used to transfer artificial chromosomes carrying the CFTR gene locus 20 or a pCMV-CFTR plasmid, 10,21 respectively, to cell lines in vitro.…”

Section: Bactofection Of Lung Epithelial Cellsmentioning

confidence: 99%

“…However, only a very small number of bacteria survive inside 293T cells (B0.0001% of the initial dose) after 48 h (data not shown), suggesting that a significant proportion of lux measured at this time point must have been generated by plasmid DNA reaching the nuclei of the CFTE29oÀ cells. As proof-of-principle that bactofection can occur in CFTE29oÀ cells, Fajac et al 14 recently demonstrated bacteria mediated gene transfer in this cell type using E. coli carrying a plasmid in which GFP expression was under the control of the eukaryotic CMV promoter as pCMV-EGFP is not leaky in bacteria. As this is the first study using a lux plasmid for bactofection, comparisons of absolute lux expression with other studies were not possible.…”

Section: Bactofection Of Lung Epithelial Cellsmentioning

confidence: 99%

“…However, Fajac et al 14 recently assessed uptake and intracellular trafficking of this E. coli vector into human CF tracheal/ bronchial cells, 16HBE (human bronchial epithelial) cells and explant outgrowths of non-CF bronchial tissue.…”

Section: Introductionmentioning

confidence: 99%

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Bactofection of lung epithelial cells in vitro and in vivo using a genetically modified Escherichia coli

et al. 2008

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Section: Bactofection Of Lung Epithelial Cellsmentioning

confidence: 99%

Section: Bactofection Of Lung Epithelial Cellsmentioning

confidence: 99%

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Bactofection of lung epithelial cells in vitro and in vivo using a genetically modified Escherichia coli

et al. 2008

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“…LLO causes the breakdown of lysosomal membranes after phagocytosis. 14,15 L. monocytogenes, which is capable of surviving inside a mammalian cell, can also be used for bactofection, but its pathological effects have to be eliminated. 16 Gene ply118 encoding a phage endopeptidase (or lysin) is responsible for bacterial lysis and subsequently for the plasmid release.…”

Section: Bacterial Strainsmentioning

confidence: 99%

“…Although most studies using bacteria for various gene therapy-related procedures are focusing on the treatment of cancer, expectations are put into the usage of prokaryotes in other clinical entities like cystic fibrosis 14,37 or ischemic diseases. 13 …”

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confidence: 99%

Bacteria in gene therapy: bactofection versus alternative gene therapy

et al. 2005

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Recent advances in gene therapy can be attributed to improvements of gene delivery vectors. New viral and nonviral transport vehicles that considerably increase the efficiency of transfection have been prepared. However, these vectors still have many disadvantages that are difficult to overcome, thus, a new approach is needed. The approach of bacterial delivery could in the future be important for gene therapy applications. In this article we try to summarize the most important modifications that are used for the preparation of applied strains, difficulties that are related with bacterial gene delivery and the current use of bactofection in animal experiments and clinical trials. Important differences to the alternative gene therapy (AGT) are discussed. AGT resembles bacteria-mediated protein delivery, as the therapeutical proteins are produced not by host cells but by the bacteria in situ and the expression can be regulated exogenously. Although the procedure of bacterial gene delivery is far from being definitely solved, bactofection remains a promising technique for transfection in human gene therapy.

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Engineering of Escherichia coli for targeted delivery of transgenes to HER2/neu‐positive tumor cells

Chang

Cheng

Chen

et al. 2011

Biotech & Bioengineering

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Targeting of non-phagocytic tumor cells and prompt release of gene cargos upon entry into tumors are two limiting steps in the bacterial gene delivery path. To tackle these problems, the non-pathogenic Escherichia coli strain BL21(DE3) was engineered to display the anti-HER2/neu affibody on the surface. After co-incubation with tumor cells for 3 h, the anti-HER2/neu affibody-presenting E. coli strain was selectively internalized into HER2/neu-positive SKBR-3 cells. The invasion efficiency reached as high as 30%. Furthermore, the bacteria were equipped with the phage ϕX174 lysin gene E-mediated autolysis system. Carrying the transgene (e.g., eukaryotic green fluorescent protein, GFP), the tumor-targeting bacteria were subjected to the thermal shock to trigger the autolysis system upon entry into HER2/neu-positive cells. Flow cytometric analysis revealed that 3% of infected cells expressed GFP 24 h post thermal induction. Overall, the results show a promise of the proposed approach for developing bacteria as a delivery carrier.

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Recombinant Escherichia coli as a gene delivery vector into airway epithelial cells

Cited by 29 publications

References 18 publications

Bactofection of lung epithelial cells in vitro and in vivo using a genetically modified Escherichia coli

Bactofection of lung epithelial cells in vitro and in vivo using a genetically modified Escherichia coli

Bacteria in gene therapy: bactofection versus alternative gene therapy

Engineering of Escherichia coli for targeted delivery of transgenes to HER2/neu‐positive tumor cells

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