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Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene

BACKGROUND: Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body. AIM OF WORK: Th...

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Autores principales: Sabet, Salwa, George, Marina A., El-Shorbagy, Haidan M., Bassiony, Heba, Farroh, Khaled Y., Youssef, Tareq, Salaheldin, Taher A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5528829/
https://www.ncbi.nlm.nih.gov/pubmed/28746382
http://dx.doi.org/10.1371/journal.pone.0181723
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author Sabet, Salwa
George, Marina A.
El-Shorbagy, Haidan M.
Bassiony, Heba
Farroh, Khaled Y.
Youssef, Tareq
Salaheldin, Taher A.
author_facet Sabet, Salwa
George, Marina A.
El-Shorbagy, Haidan M.
Bassiony, Heba
Farroh, Khaled Y.
Youssef, Tareq
Salaheldin, Taher A.
author_sort Sabet, Salwa
collection PubMed
description BACKGROUND: Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body. AIM OF WORK: The present study aimed to develop stable Gel.NPs conjugated with nonstructural protein 2 (NS2) gene of Hepatitis C Virus genotype 4a (HCV4a) as a safe and an efficient vaccine delivery system. METHODS AND RESULTS: Gel.NPs were synthesized and characterized (size: 150±2 nm and zeta potential +17.6 mv). NS2 gene was successfully cloned and expressed into E. coli M15 using pQE-30 vector. Antigenicity of the recombinant NS2 protein was confirmed by Western blotting to verify the efficiency of NS2 as a possible vaccine. Then NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM). Interestingly, the transformation of the conjugated NS2/Gel.NPs complex into E. coli DH5-α was 50% more efficient than transformation with the gene alone. In addition, conjugated NS2/Gel.NPs with ratio 1:100 (w/w) showed higher transformation efficiency into E. coli DH5-α than the other ratios (1:50 and 2:50). CONCLUSION: Gel.NPs effectively enhanced the gene delivery in bacterial cells without affecting the structure of NS2 gene and could be used as a safe, easy, rapid, cost-effective and non-viral vaccine delivery system for HCV.
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spelling pubmed-55288292017-08-07 Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene Sabet, Salwa George, Marina A. El-Shorbagy, Haidan M. Bassiony, Heba Farroh, Khaled Y. Youssef, Tareq Salaheldin, Taher A. PLoS One Research Article BACKGROUND: Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body. AIM OF WORK: The present study aimed to develop stable Gel.NPs conjugated with nonstructural protein 2 (NS2) gene of Hepatitis C Virus genotype 4a (HCV4a) as a safe and an efficient vaccine delivery system. METHODS AND RESULTS: Gel.NPs were synthesized and characterized (size: 150±2 nm and zeta potential +17.6 mv). NS2 gene was successfully cloned and expressed into E. coli M15 using pQE-30 vector. Antigenicity of the recombinant NS2 protein was confirmed by Western blotting to verify the efficiency of NS2 as a possible vaccine. Then NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM). Interestingly, the transformation of the conjugated NS2/Gel.NPs complex into E. coli DH5-α was 50% more efficient than transformation with the gene alone. In addition, conjugated NS2/Gel.NPs with ratio 1:100 (w/w) showed higher transformation efficiency into E. coli DH5-α than the other ratios (1:50 and 2:50). CONCLUSION: Gel.NPs effectively enhanced the gene delivery in bacterial cells without affecting the structure of NS2 gene and could be used as a safe, easy, rapid, cost-effective and non-viral vaccine delivery system for HCV. Public Library of Science 2017-07-26 /pmc/articles/PMC5528829/ /pubmed/28746382 http://dx.doi.org/10.1371/journal.pone.0181723 Text en © 2017 Sabet et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sabet, Salwa
George, Marina A.
El-Shorbagy, Haidan M.
Bassiony, Heba
Farroh, Khaled Y.
Youssef, Tareq
Salaheldin, Taher A.
Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title_full Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title_fullStr Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title_full_unstemmed Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title_short Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene
title_sort gelatin nanoparticles enhance delivery of hepatitis c virus recombinant ns2 gene
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5528829/
https://www.ncbi.nlm.nih.gov/pubmed/28746382
http://dx.doi.org/10.1371/journal.pone.0181723
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