Document Detail


A novel plasmid for delivering genes into mammalian cells with noninvasive food and commensal lactic acid bacteria.
MedLine Citation:
PMID:  20832422     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Using food and commensal lactic acid bacteria (LAB) as vehicles for DNA delivery into epithelial cells is a new strategy for vaccine delivery or gene therapy. However, present methods for DNA delivery with LAB have suffered low efficiency. Our goal was to develop a new system to deliver DNA into epithelial cells with high efficiency using food and commensal LAB. An Escherichia coli-LAB shuttle plasmid, pLKV1, for DNA delivery into eukaryotic cells was constructed. Two reporter plasmids with green and red fluorescent protein genes were also constructed to monitor the uptake of protein and DNA, respectively. Bacteria delivering these reporter plasmids into Caco-2 cells were monitored by fluorescence microscopy. Several methods that weaken the bacterial cell wall prior to co-culture with Caco-2 cells were evaluated for their role in the improvement of gene transfer efficiency. Treating Streptococcus gordonii with penicillin and lysozyme greatly increased its rate of gene delivery to mammalian cells compared to untreated control bacteria, while glycine pretreatment promoted the highest gene transfer rate for Lactococcus lactis. Uptake of green fluorescent bacteria by Caco-2 cells showed that the cell wall-weakening treatment promoted the internalization of the noninvasive bacteria into Caco-2 cells. In conclusion, we have developed a noninvasive system using LAB as a vehicle for vaccine delivery or gene therapy, and tested this system in vitro with Caco-2 cells.
Authors:
Lin Tao; Sylvia I Pavlova; Xin Ji; Ling Jin; Gregory Spear
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-09-09
Journal Detail:
Title:  Plasmid     Volume:  65     ISSN:  1095-9890     ISO Abbreviation:  Plasmid     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-01-04     Completed Date:  2011-04-15     Revised Date:  2013-06-10    
Medline Journal Info:
Nlm Unique ID:  7802221     Medline TA:  Plasmid     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8-14     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Inc. All rights reserved.
Affiliation:
Department of Oral Biology, College of Dentistry, University of Illinois at Chicago, 801 South Paulina Street, Chicago, IL 60612, USA. ltao@uic.edu
Data Bank Information
Bank Name/Acc. No.:
GENBANK/HM569775
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MeSH Terms
Descriptor/Qualifier:
Bacteria / cytology,  metabolism*
Caco-2 Cells
Cell Wall / metabolism
Escherichia coli / cytology,  metabolism
Food Microbiology*
Genes, Reporter / genetics
Green Fluorescent Proteins / metabolism
Humans
Lactic Acid / metabolism*
Lactococcus / cytology,  metabolism
Luminescent Proteins / metabolism
Molecular Sequence Data
Plasmids / genetics*
Streptococcus / cytology,  metabolism
Transfection / methods*
Grant Support
ID/Acronym/Agency:
AI066709/AI/NIAID NIH HHS; P30 AI082151/AI/NIAID NIH HHS; R21 AI050491-02/AI/NIAID NIH HHS; U01 AI066709-05/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 0/red fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 50-21-5/Lactic Acid
Comments/Corrections

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