A new Broad range plasmid for DNA delivery in eukaryotic cells using lactic acid bacteria: In vitro and In vivo assays - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Molecular Therapy - Methods and Clinical Development Année : 2017

A new Broad range plasmid for DNA delivery in eukaryotic cells using lactic acid bacteria: In vitro and In vivo assays

Résumé

Lactococcus lactis is well documented as a promising candidate for development of novel oral live vaccines. It has been broadly engineered for heterologous expression, as well as for plasmid expression vector delivery, directly inside eukaryotic cells, for DNA vaccine, or as therapeutic vehicle. This work describes the characteristics of a new plasmid, pExu (extra chromosomal unit), for DNA delivery using L. lactis and evaluates its functionality both by in vitro and in vivo assays. This plasmid exhibits the following features: (1) a theta origin of replication and (2) an expression cassette containing a multiple cloning site and a eukaryotic promoter, the cytomegalovirus (pCMV). The functionality of pExu: egfp was evaluated by fluorescence microscopy. The L. lactis MG1363 (pExu: egfp) strains were administered by gavage to Balb/C mice and the eGFP expression was monitored by fluorescence microscopy. The pExu vector has demonstrated an excellent stability either in L. lactis or in Escherichia coli. The eGFP expression at different times in in vitro assay showed that 15.8% of CHO cells were able to express the protein after transfection. The enterocytes of mice showed the expression of eGFP protein. Thus, L. lactis carrying the pExu is a good candidate to deliver genes into eukaryotic cells.
Fichier principal
Vignette du fichier
2017_Mancha-Agresti_Molecular therapy. Methods and clinical development_1.pdf (1.87 Mo) Télécharger le fichier
Origine : Fichiers éditeurs autorisés sur une archive ouverte
Loading...

Dates et versions

hal-01604769 , version 1 (26-05-2020)

Licence

Paternité - Pas d'utilisation commerciale - Pas de modification

Identifiants

Citer

Pamela Mancha-Agresti, Mariana Martins Drumond, Fillipe Luiz Rosa Do Carmo, Monica Morais Santos, Janete Soares Coelho dos Santos, et al.. A new Broad range plasmid for DNA delivery in eukaryotic cells using lactic acid bacteria: In vitro and In vivo assays. Molecular Therapy - Methods and Clinical Development, 2017, 4, pp.1-9. ⟨10.1016/j.omtm.2016.12.005⟩. ⟨hal-01604769⟩

Collections

ALIMH
30 Consultations
43 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More