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Para amino benzoic acid-derived self-assembled biocompatible nanoparticles for efficient delivery of siRNA.

Reddy TL, Krishnarao PS, Rao GK, Bhimireddy E, Venkateswarlu P, Mohapatra DK, Yadav JS, Bhadra U, Bhadra MP - Int J Nanomedicine (2015)

Bottom Line: A number of diseases can result from abnormal gene expression.Our findings indicated high gene transfection efficiency.These biocompatible nanoparticles allow targeted delivery of siRNA, providing an efficient vehicle for gene delivery.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chemical Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad, India ; Academy of Scientific and Innovative Research, New Delhi, India.

ABSTRACT
A number of diseases can result from abnormal gene expression. One of the approaches for treating such diseases is gene therapy to inhibit expression of a particular gene in a specific cell population by RNA interference. Use of efficient delivery vehicles increases the safety and success of gene therapy. Here we report the development of functionalized biocompatible fluorescent nanoparticles from para amino benzoic acid nanoparticles for efficient delivery of short interfering RNA (siRNA). These nanoparticles were non-toxic and did not interfere with progression of the cell cycle. The intrinsic fluorescent nature of these nanoparticles allows easy tracking and an opportunity for diagnostic applications. Human Bcl-2 siRNA was complexed with these nanoparticles to inhibit expression in cells at both the transcriptional and translational levels. Our findings indicated high gene transfection efficiency. These biocompatible nanoparticles allow targeted delivery of siRNA, providing an efficient vehicle for gene delivery.

No MeSH data available.


Related in: MedlinePlus

Scanning electron microscopic images of NPs and confocal images of NP-siRNA complex (nanoplex) 12 hours post transfection.Notes: NPs exhibits intrinsic fluorescence (green) and siRNA was 5′-labelled with Cy3. NPs were complexed with siRNA at a ratio of 20:1 (w/w). Images shown represent each separate channel, with NPs in green, siRNA in red, and the merged images shown on the top right. Scale 250 nm (SEM), 20 μm (cells).Abbreviations: NPs, nanoparticles; siRNA, short interfering RNA; SEM, scanning electron microscopy.
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f3-ijn-10-6411: Scanning electron microscopic images of NPs and confocal images of NP-siRNA complex (nanoplex) 12 hours post transfection.Notes: NPs exhibits intrinsic fluorescence (green) and siRNA was 5′-labelled with Cy3. NPs were complexed with siRNA at a ratio of 20:1 (w/w). Images shown represent each separate channel, with NPs in green, siRNA in red, and the merged images shown on the top right. Scale 250 nm (SEM), 20 μm (cells).Abbreviations: NPs, nanoparticles; siRNA, short interfering RNA; SEM, scanning electron microscopy.

Mentions: PABA molecules were synthesized starting with PABA using the afore-mentioned methods, and characterized using 1H-NMR and 13C-NMR (Figures S4 and S5), mass spectroscopy, and infrared spectroscopy. The synthesized compounds were then allowed to self-assemble. The PABA nanomaterials thus obtained from compound (8a–f) were named G-10, G-12, G-14, G-16, G-18, and G-18u respectively, based on the length of the side chains and unsaturated moieties coupled during synthesis. To characterize the physiochemical properties of the functionalized NPs (G10, G12, G14, G16, G18, and G18U) and to examine their structural and spectral properties, freshly prepared NPs were dispersed in water. All the NPs were readily soluble and stable in water. An important consideration in siRNA delivery is the overall size of the NP-siRNA complexes. We performed scanning electron microscopy and DLS to observe the morphology and size of the NPs and when complexed with siRNA. The NPs and NP-siRNA complexes dispersed readily and formed well defined structures (Figure 3). DLS studies confirmed the NPs to have an overall size range of 100–200 nm (Table 1). Confocal microscopic images showed that the six nanostructures containing a guanidine modification and bearing saturated or unsaturated acid side chains exhibited intrinsic green fluorescence.


Para amino benzoic acid-derived self-assembled biocompatible nanoparticles for efficient delivery of siRNA.

Reddy TL, Krishnarao PS, Rao GK, Bhimireddy E, Venkateswarlu P, Mohapatra DK, Yadav JS, Bhadra U, Bhadra MP - Int J Nanomedicine (2015)

Scanning electron microscopic images of NPs and confocal images of NP-siRNA complex (nanoplex) 12 hours post transfection.Notes: NPs exhibits intrinsic fluorescence (green) and siRNA was 5′-labelled with Cy3. NPs were complexed with siRNA at a ratio of 20:1 (w/w). Images shown represent each separate channel, with NPs in green, siRNA in red, and the merged images shown on the top right. Scale 250 nm (SEM), 20 μm (cells).Abbreviations: NPs, nanoparticles; siRNA, short interfering RNA; SEM, scanning electron microscopy.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608593&req=5

f3-ijn-10-6411: Scanning electron microscopic images of NPs and confocal images of NP-siRNA complex (nanoplex) 12 hours post transfection.Notes: NPs exhibits intrinsic fluorescence (green) and siRNA was 5′-labelled with Cy3. NPs were complexed with siRNA at a ratio of 20:1 (w/w). Images shown represent each separate channel, with NPs in green, siRNA in red, and the merged images shown on the top right. Scale 250 nm (SEM), 20 μm (cells).Abbreviations: NPs, nanoparticles; siRNA, short interfering RNA; SEM, scanning electron microscopy.
Mentions: PABA molecules were synthesized starting with PABA using the afore-mentioned methods, and characterized using 1H-NMR and 13C-NMR (Figures S4 and S5), mass spectroscopy, and infrared spectroscopy. The synthesized compounds were then allowed to self-assemble. The PABA nanomaterials thus obtained from compound (8a–f) were named G-10, G-12, G-14, G-16, G-18, and G-18u respectively, based on the length of the side chains and unsaturated moieties coupled during synthesis. To characterize the physiochemical properties of the functionalized NPs (G10, G12, G14, G16, G18, and G18U) and to examine their structural and spectral properties, freshly prepared NPs were dispersed in water. All the NPs were readily soluble and stable in water. An important consideration in siRNA delivery is the overall size of the NP-siRNA complexes. We performed scanning electron microscopy and DLS to observe the morphology and size of the NPs and when complexed with siRNA. The NPs and NP-siRNA complexes dispersed readily and formed well defined structures (Figure 3). DLS studies confirmed the NPs to have an overall size range of 100–200 nm (Table 1). Confocal microscopic images showed that the six nanostructures containing a guanidine modification and bearing saturated or unsaturated acid side chains exhibited intrinsic green fluorescence.

Bottom Line: A number of diseases can result from abnormal gene expression.Our findings indicated high gene transfection efficiency.These biocompatible nanoparticles allow targeted delivery of siRNA, providing an efficient vehicle for gene delivery.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chemical Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad, India ; Academy of Scientific and Innovative Research, New Delhi, India.

ABSTRACT
A number of diseases can result from abnormal gene expression. One of the approaches for treating such diseases is gene therapy to inhibit expression of a particular gene in a specific cell population by RNA interference. Use of efficient delivery vehicles increases the safety and success of gene therapy. Here we report the development of functionalized biocompatible fluorescent nanoparticles from para amino benzoic acid nanoparticles for efficient delivery of short interfering RNA (siRNA). These nanoparticles were non-toxic and did not interfere with progression of the cell cycle. The intrinsic fluorescent nature of these nanoparticles allows easy tracking and an opportunity for diagnostic applications. Human Bcl-2 siRNA was complexed with these nanoparticles to inhibit expression in cells at both the transcriptional and translational levels. Our findings indicated high gene transfection efficiency. These biocompatible nanoparticles allow targeted delivery of siRNA, providing an efficient vehicle for gene delivery.

No MeSH data available.


Related in: MedlinePlus