A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays
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. Thi...
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| Format: | Article |
| Language: | English |
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Elsevier
2017-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050116301383 |
| _version_ | 1811236727204872192 |
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| author | Pamela Mancha-Agresti Mariana Martins Drumond Fillipe Luiz Rosa do Carmo Monica Morais Santos Janete Soares Coelho dos Santos Franco Venanzi Jean-Marc Chatel Sophie Yvette Leclercq Vasco Azevedo |
| author_facet | Pamela Mancha-Agresti Mariana Martins Drumond Fillipe Luiz Rosa do Carmo Monica Morais Santos Janete Soares Coelho dos Santos Franco Venanzi Jean-Marc Chatel Sophie Yvette Leclercq Vasco Azevedo |
| author_sort | Pamela Mancha-Agresti |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T12:13:00Z |
| format | Article |
| id | doaj.art-f6dfbd21a1a844aea4dac4fdbe09819e |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-04-12T12:13:00Z |
| publishDate | 2017-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-f6dfbd21a1a844aea4dac4fdbe09819e2022-12-22T03:33:31ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012017-03-014C839110.1016/j.omtm.2016.12.005A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo AssaysPamela Mancha-Agresti0Mariana Martins Drumond1Fillipe Luiz Rosa do Carmo2Monica Morais Santos3Janete Soares Coelho dos Santos4Franco Venanzi5Jean-Marc Chatel6Sophie Yvette Leclercq7Vasco Azevedo8Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB/UFMG), 31270-901 Belo Horizonte, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB/UFMG), 31270-901 Belo Horizonte, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB/UFMG), 31270-901 Belo Horizonte, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB/UFMG), 31270-901 Belo Horizonte, BrazilLaboratório de Inovação Biotecnológica, Fundação Ezequiel Dias (FUNED), Belo Horizonte, 30510-010 Minas Gerais, BrazilSchool of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino, ItalyMicalis Institute, INRA, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, FranceLaboratório de Inovação Biotecnológica, Fundação Ezequiel Dias (FUNED), Belo Horizonte, 30510-010 Minas Gerais, BrazilInstituto de Ciências Biológicas, Universidade Federal de Minas Gerais (ICB/UFMG), 31270-901 Belo Horizonte, BrazilLactococcus 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.http://www.sciencedirect.com/science/article/pii/S2329050116301383lactic acid bacteriaLactococcus lactisDNA mucosal deliveringshuttle plasmid |
| spellingShingle | Pamela Mancha-Agresti Mariana Martins Drumond Fillipe Luiz Rosa do Carmo Monica Morais Santos Janete Soares Coelho dos Santos Franco Venanzi Jean-Marc Chatel Sophie Yvette Leclercq Vasco Azevedo A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays Molecular Therapy: Methods & Clinical Development lactic acid bacteria Lactococcus lactis DNA mucosal delivering shuttle plasmid |
| title | A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays |
| title_full | A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays |
| title_fullStr | A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays |
| title_full_unstemmed | A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays |
| title_short | A New Broad Range Plasmid for DNA Delivery in Eukaryotic Cells Using Lactic Acid Bacteria: In Vitro and In Vivo Assays |
| title_sort | new broad range plasmid for dna delivery in eukaryotic cells using lactic acid bacteria in vitro and in vivo assays |
| topic | lactic acid bacteria Lactococcus lactis DNA mucosal delivering shuttle plasmid |
| url | http://www.sciencedirect.com/science/article/pii/S2329050116301383 |
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