Staff View: Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression

Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression

The delivery of a full plasmid, encoding the green fluorescent protein gene into African monkey kidney (Vero3) cells, was successfully achieved using nanobiocomposites based on layered double hydroxides. This demonstrated the potential of using the system as an alternative DNA delivery vector. Inte...

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Main Authors:	Masarudin, Mas Jaffri, Yusoff, Khatijah, Abdul Rahim, Raha, Hussein, Mohd. Zobir
Format:	Article
Language:	English English
Published:	IOI Publishing 2009
Subjects:	Plasmids - Genetics DNA
Online Access:	http://psasir.upm.edu.my/id/eprint/16887/1/Successful%20transfer%20of%20plasmid%20DNA%20into%20in%20vitro%20cells%20transfected%20with%20an%20inorganic%20plasmidMg.pdf
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id	oai:psasir.upm.edu.my:16887
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spelling	oai:psasir.upm.edu.my:16887 http://psasir.upm.edu.my/id/eprint/16887/ Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression Masarudin, Mas Jaffri Yusoff, Khatijah Abdul Rahim, Raha Hussein, Mohd. Zobir The delivery of a full plasmid, encoding the green fluorescent protein gene into African monkey kidney (Vero3) cells, was successfully achieved using nanobiocomposites based on layered double hydroxides. This demonstrated the potential of using the system as an alternative DNA delivery vector. Intercalation of the circular plasmid DNA, pEGFP-N2, into Mg/Al-NO−3 layered double hydroxides (LDH) was accomplished through anion exchange routes to form the nanobiocomposite material. The host was previously synthesized at the Mg2+ to Al3+ molar ratio Ri = 2 and subsequently intercalated with plasmid DNA. Size expansion of the interlamellae host from 8.8 ° A in LDH to 42 °A was observed in the resulting nanobiocomposite,indicating stable hybridization of the plasmid DNA. The powder x-ray diffraction (PXRD)results, supplemented with Fourier-transform infrared (FTIR) spectroscopy, compositional and electrophoresis studies confirmed the encapsulation episode of the biomaterial. In order to elucidate the use of this resulting nanobiocomposite as a delivery vector, an MTT assay was performed to determine any cytotoxic effects of the host towards cells. The intercalated pEGFP-N2 anion was later successfully recovered through acidification with HNO3 after treatment with DNA-degrading enzymes, thus also showing the ability of the LDH host to protect the intercalated biomaterial from degradation. Cell transfection studies on Vero3 cells were then performed, where cells transfected with the nanobiocomposite exhibited fluorescence as early as 12 h post-treatment compared to naked delivery of the plasmid itself. IOI Publishing 2009 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/16887/1/Successful%20transfer%20of%20plasmid%20DNA%20into%20in%20vitro%20cells%20transfected%20with%20an%20inorganic%20plasmidMg.pdf Masarudin, Mas Jaffri and Yusoff, Khatijah and Abdul Rahim, Raha and Hussein, Mohd. Zobir (2009) Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression. Nanotechnology, 20. pp. 1-11. ISSN 1361-6528 Plasmids - Genetics DNA doi:10.1088/0957-4484/20/4/045602 English
institution	UPM IR
collection	UPM IR
language	English English
topic	Plasmids - Genetics DNA
spellingShingle	Plasmids - Genetics DNA Masarudin, Mas Jaffri Yusoff, Khatijah Abdul Rahim, Raha Hussein, Mohd. Zobir Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
description	The delivery of a full plasmid, encoding the green fluorescent protein gene into African monkey kidney (Vero3) cells, was successfully achieved using nanobiocomposites based on layered double hydroxides. This demonstrated the potential of using the system as an alternative DNA delivery vector. Intercalation of the circular plasmid DNA, pEGFP-N2, into Mg/Al-NO−3 layered double hydroxides (LDH) was accomplished through anion exchange routes to form the nanobiocomposite material. The host was previously synthesized at the Mg2+ to Al3+ molar ratio Ri = 2 and subsequently intercalated with plasmid DNA. Size expansion of the interlamellae host from 8.8 ° A in LDH to 42 °A was observed in the resulting nanobiocomposite,indicating stable hybridization of the plasmid DNA. The powder x-ray diffraction (PXRD)results, supplemented with Fourier-transform infrared (FTIR) spectroscopy, compositional and electrophoresis studies confirmed the encapsulation episode of the biomaterial. In order to elucidate the use of this resulting nanobiocomposite as a delivery vector, an MTT assay was performed to determine any cytotoxic effects of the host towards cells. The intercalated pEGFP-N2 anion was later successfully recovered through acidification with HNO3 after treatment with DNA-degrading enzymes, thus also showing the ability of the LDH host to protect the intercalated biomaterial from degradation. Cell transfection studies on Vero3 cells were then performed, where cells transfected with the nanobiocomposite exhibited fluorescence as early as 12 h post-treatment compared to naked delivery of the plasmid itself.
format	Article
author	Masarudin, Mas Jaffri Yusoff, Khatijah Abdul Rahim, Raha Hussein, Mohd. Zobir
author_facet	Masarudin, Mas Jaffri Yusoff, Khatijah Abdul Rahim, Raha Hussein, Mohd. Zobir
author_sort	Masarudin, Mas Jaffri
title	Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
title_short	Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
title_full	Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
title_fullStr	Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
title_full_unstemmed	Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression
title_sort	successful transfer of plasmid dna into in vitro cells transfected with an inorganic plasmidmg/al-ldh nanobiocomposite material as a vector for gene expression
publisher	IOI Publishing
publishDate	2009
url	http://psasir.upm.edu.my/id/eprint/16887/1/Successful%20transfer%20of%20plasmid%20DNA%20into%20in%20vitro%20cells%20transfected%20with%20an%20inorganic%20plasmidMg.pdf
_version_	1819294077417422848
score	13.4562235

Successful transfer of plasmid DNA into in vitro cells transfected with an inorganic plasmidMg/Al-LDH nanobiocomposite material as a vector for gene expression

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