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A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid
Bacteria: In Vitro and In Vivo Assays
#MMPMID28344994
Mancha-Agresti P
; Drumond MM
; Carmo FL
; Santos MM
; Santos JS
; Venanzi F
; Chatel JM
; Leclercq SY
; Azevedo V
Mol Ther Methods Clin Dev
2017[Mar]; 4
(?): 83-91
PMID28344994
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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.
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