Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery
PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccine, polyamidoamine (PAMAM) dendrimers conjugated with HIV transactivator of transcription (TAT) was developed. METHODS: First, the plasmid DNA (pIRES-H5/GFP) nanoparticle was formulated using PAMAM d...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Canadian Society for Pharmaceutical Sciences
2016
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| Online Access: | http://psasir.upm.edu.my/id/eprint/55505/ http://psasir.upm.edu.my/id/eprint/55505/1/Development%20of%20Tat-Conjugated%20Dendrimer%20for%20Transdermal%20DNA%20Vaccine%20Delivery..pdf |
| _version_ | 1848852821109112832 |
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| author | Bahadoran, Azadeh Moeini, Hassan Bejo, Mohd Hair Hussein, Mohd Zobir Omar, Abdul Rahman |
| author_facet | Bahadoran, Azadeh Moeini, Hassan Bejo, Mohd Hair Hussein, Mohd Zobir Omar, Abdul Rahman |
| author_sort | Bahadoran, Azadeh |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | PURPOSE:
In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccine, polyamidoamine (PAMAM) dendrimers conjugated with HIV transactivator of transcription (TAT) was developed.
METHODS:
First, the plasmid DNA (pIRES-H5/GFP) nanoparticle was formulated using PAMAM dendrimer and TAT peptide and then characterized for surface charge, particle size, DNA encapsulation and protection of the pIRES-H5/GFP DNA plasmid to enzymatic digestion. Subsequently, the potency of the TAT-conjugated dendrimer for gene delivery was evaluated through in vitro transfection into Vero cells followed by gene expression analysis including western blotting, fluorescent microscopy and PCR. The effect of the TAT peptide on cellular uptake of DNA vaccine was studied by qRT-PCR and flow cytometry. Finally, the ability of TAT-conjugated PAMAM dendrimer for transdermal delivery of the DNA plasmid was assessed through artificial membranes followed by qRT-PCR and flow cytometry.
RESULTS:
TAT-conjugated PAMAM dendrimer showed the ability to form a compact and nanometre-sized polyplexes with the plasmid DNA, having the size range of 105 to 115 nm and a positive charge of +42 to +45 mV over the N/P ratio of 6:1(+/-). In vitro transfection analysis into Vero cells confirms the high potency of TAT-conjugated PAMAM dendrimer to enhance the cellular uptake of DNA vaccine. The permeability value assay through artificial membranes reveals that TAT-conjugated PAMAM has more capacity for transdermal delivery of the DNA compared to unmodified PAMAM dendrimer (P<0.05).
CONCLUSIONS:
The findings of this study suggest that TAT-conjugated PAMAM dendrimer is a promising non-viral vector for transdermal use.This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page. |
| first_indexed | 2025-11-15T10:44:10Z |
| format | Article |
| id | upm-55505 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:44:10Z |
| publishDate | 2016 |
| publisher | Canadian Society for Pharmaceutical Sciences |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-555052017-09-13T10:09:16Z http://psasir.upm.edu.my/id/eprint/55505/ Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery Bahadoran, Azadeh Moeini, Hassan Bejo, Mohd Hair Hussein, Mohd Zobir Omar, Abdul Rahman PURPOSE: In order to enhance cellular uptake and to facilitate transdermal delivery of DNA vaccine, polyamidoamine (PAMAM) dendrimers conjugated with HIV transactivator of transcription (TAT) was developed. METHODS: First, the plasmid DNA (pIRES-H5/GFP) nanoparticle was formulated using PAMAM dendrimer and TAT peptide and then characterized for surface charge, particle size, DNA encapsulation and protection of the pIRES-H5/GFP DNA plasmid to enzymatic digestion. Subsequently, the potency of the TAT-conjugated dendrimer for gene delivery was evaluated through in vitro transfection into Vero cells followed by gene expression analysis including western blotting, fluorescent microscopy and PCR. The effect of the TAT peptide on cellular uptake of DNA vaccine was studied by qRT-PCR and flow cytometry. Finally, the ability of TAT-conjugated PAMAM dendrimer for transdermal delivery of the DNA plasmid was assessed through artificial membranes followed by qRT-PCR and flow cytometry. RESULTS: TAT-conjugated PAMAM dendrimer showed the ability to form a compact and nanometre-sized polyplexes with the plasmid DNA, having the size range of 105 to 115 nm and a positive charge of +42 to +45 mV over the N/P ratio of 6:1(+/-). In vitro transfection analysis into Vero cells confirms the high potency of TAT-conjugated PAMAM dendrimer to enhance the cellular uptake of DNA vaccine. The permeability value assay through artificial membranes reveals that TAT-conjugated PAMAM has more capacity for transdermal delivery of the DNA compared to unmodified PAMAM dendrimer (P<0.05). CONCLUSIONS: The findings of this study suggest that TAT-conjugated PAMAM dendrimer is a promising non-viral vector for transdermal use.This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page. Canadian Society for Pharmaceutical Sciences 2016 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/55505/1/Development%20of%20Tat-Conjugated%20Dendrimer%20for%20Transdermal%20DNA%20Vaccine%20Delivery..pdf Bahadoran, Azadeh and Moeini, Hassan and Bejo, Mohd Hair and Hussein, Mohd Zobir and Omar, Abdul Rahman (2016) Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery. Journal of Pharmacy and Pharmaceutical Sciences, 19 (3). pp. 325-338. ISSN 1482-1826 http://www.ualberta.ca/~csps/Journals/JPPS.htm 10.18433/J3G31Q |
| spellingShingle | Bahadoran, Azadeh Moeini, Hassan Bejo, Mohd Hair Hussein, Mohd Zobir Omar, Abdul Rahman Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title | Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title_full | Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title_fullStr | Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title_full_unstemmed | Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title_short | Development of tat - conjugated dendrimer for transdermal DNA vaccine delivery |
| title_sort | development of tat - conjugated dendrimer for transdermal dna vaccine delivery |
| url | http://psasir.upm.edu.my/id/eprint/55505/ http://psasir.upm.edu.my/id/eprint/55505/ http://psasir.upm.edu.my/id/eprint/55505/ http://psasir.upm.edu.my/id/eprint/55505/1/Development%20of%20Tat-Conjugated%20Dendrimer%20for%20Transdermal%20DNA%20Vaccine%20Delivery..pdf |