title: Construction of a new shuttle vector for DNA delivery into mammalian cells using non-invasive Lactococcus lactis.
abstract: Use of food grade Lactococcus lactis (L. lactis) is fast emerging as a safe alternative for delivery of DNA vaccine. To attain efficient DNA delivery, L. lactis, a non-invasive bacterium is converted to invasive strain either by expressing proteins like Internalin A (InlA) or Fibronectin binding protein A (FnBPA) or through chemical treatments. However the safety status of invasive L. lactis is questionable. In the present report, we have shown that non-invasive L. lactis efficiently delivered the newly constructed reporter plasmid pPERDBY to mammalian cells without any chemical enhancers. The salient features of the vector are; I) Ability to replicate in two different hosts; Escherichia coli (E. coli) and Lactic Acid Bacteria (LAB), II) One of the smallest reporter plasmid for DNA vaccine, III) Enhanced Green Fluorescence Protein (EGFP) linked to Multiple Cloning Site (MCS), IV) Immunostimulatory CpG motifs functioning as an adjuvant. Expression of EGFP in pPERDBY transfected CHO-K1 and Caco-2 cells demonstrates its functionality. Non-invasive r-L. lactis was found efficient in delivering pPERDBY to Caco-2 cells. The in vitro data presented in this article supports the hypothesis that in the absence of invasive proteins or relevant chemical treatment, L. lactis was found efficient in delivering DNA to mammalian cells.
authors.affiliation: Department of Cell and Molecular Biology, B. V. Patel Pharmaceutical Education and Research Development (PERD) Centre, Ahmedabad 380054, Gujarat, India. Electronic address: bjyajnik3@gmail.com.
authors.forename: Bhrugu
authors.initials: B
authors.lastname: Yagnik
authors.affiliation: Sardar Patel University, Vallabh Vidhyanagar 388120, Gujarat, India. Electronic address: hpadh@yahoo.com.
authors.forename: Harish
authors.initials: H
authors.lastname: Padh
authors.affiliation: Department of Cell and Molecular Biology, B. V. Patel Pharmaceutical Education and Research Development (PERD) Centre, Ahmedabad 380054, Gujarat, India. Electronic address: preetindesai79@gmail.com.
authors.forename: Priti
authors.initials: P
authors.lastname: Desai
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics; D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D018938:Caco-2 Cells; D003412:Cricetulus; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D013294:Lactococcus lactis / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D010957:Plasmids; D014162:Transfection
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D009838:Oligodeoxyribonucleotides; D004247:DNA
keywords: DNA vaccine; EGFP; Lactococcus lactis; Non-invasive; Plasmid; Vaccine delivery