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Nanotherapeutics Using an HIV-1 Poly A and Transactivator of the HIV-1 LTR-(TAR-) Specific siRNA.

Mahajan SD, Aalinkeel R, Reynolds JL, Nair B, Sykes DE, Law WC, Ding H, Bergey EJ, Prasad PN, Schwartz SA - Patholog Res Int (2011)

Bottom Line: We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication.Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period.HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

View Article: PubMed Central - PubMed

Affiliation: Division of Allergy, Immunology, and Rheumatology, Department of Medicine, University at Buffalo, The State University of New York, 640 Ellicott Street, Room 444 Innovation Center, Buffalo, NY 14203, USA.

ABSTRACT
HIV-1 replication can be efficiently inhibited by intracellular expression of an siRNA targeting the viral RNA. We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication. Nanotechnology holds much potential for impact in the field of HIV-1 therapeutics, and nanoparticles such as quantum rods (QRs) can be easily functionalized to incorporate siRNA forming stable nanoplexes that can be used for gene silencing. We evaluated the efficacy of the QR-si510 HIV-1 siRNA nanoplex in suppressing viral replication in the HIV-1-infected monocytic cell line THP-1 by measuring p24 antigen levels and gene expression levels of HIV-1 LTR. Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period. HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

No MeSH data available.


Related in: MedlinePlus

Confocal images of THP-1 cells treated QR-si510 siRNA nanoplexes: THP-1 cells were treated with QR-si510 siRNA nanoplexes for 12, 48, 96 hr, and up to 1 week and uptake of the nanoplexes was observed using confocal microscopy. (a) Representative local emission spectra confirms that the fluorescence signal in the THP-1 cells is localized to the QR si510 HIV-1siRNA nanoplexes, evident from the sharp emission spectra peak of 625 nm. (b) (A–J): Parallel Fluorescence and transmission images showing fluorescence (red) staining indicating QR-si510 siRNA nanoplex uptake by THP-1 cells.
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fig6: Confocal images of THP-1 cells treated QR-si510 siRNA nanoplexes: THP-1 cells were treated with QR-si510 siRNA nanoplexes for 12, 48, 96 hr, and up to 1 week and uptake of the nanoplexes was observed using confocal microscopy. (a) Representative local emission spectra confirms that the fluorescence signal in the THP-1 cells is localized to the QR si510 HIV-1siRNA nanoplexes, evident from the sharp emission spectra peak of 625 nm. (b) (A–J): Parallel Fluorescence and transmission images showing fluorescence (red) staining indicating QR-si510 siRNA nanoplex uptake by THP-1 cells.

Mentions: The cellular uptake of the QR-siRNA nanoplexes was studied using confocal microscopy. Figure 6(a) shows the confocal microscopic imaging and the local spectra of THP-1 cells treated with functional QR-si510 HIV-1siRNA nanoplexes. Confocal microscopy images were obtained with laser excitation at 405 nm. Figure 6(a) shows the representative local spectra obtained from different points in the confocal images from cells. The red labeling of THP-1 cells confirms the fluorescence signal is localized to the QR si510 HIV-1siRNA nanoplexes which is evident from the sharp emission spectra peaking around 625 nm. Additionally, it is important to note that the cellular autofluorescence which is normally located around 500 nm shows no interference with the emission from QRs. Our results show uptake of the QR-si510 HIV-1siRNA nanoplexes by the THP-1 cells at 12, 48, 96 hr, and 1 week posttransfection (Figure 6(b) (A)–(J)).


Nanotherapeutics Using an HIV-1 Poly A and Transactivator of the HIV-1 LTR-(TAR-) Specific siRNA.

Mahajan SD, Aalinkeel R, Reynolds JL, Nair B, Sykes DE, Law WC, Ding H, Bergey EJ, Prasad PN, Schwartz SA - Patholog Res Int (2011)

Confocal images of THP-1 cells treated QR-si510 siRNA nanoplexes: THP-1 cells were treated with QR-si510 siRNA nanoplexes for 12, 48, 96 hr, and up to 1 week and uptake of the nanoplexes was observed using confocal microscopy. (a) Representative local emission spectra confirms that the fluorescence signal in the THP-1 cells is localized to the QR si510 HIV-1siRNA nanoplexes, evident from the sharp emission spectra peak of 625 nm. (b) (A–J): Parallel Fluorescence and transmission images showing fluorescence (red) staining indicating QR-si510 siRNA nanoplex uptake by THP-1 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3108557&req=5

fig6: Confocal images of THP-1 cells treated QR-si510 siRNA nanoplexes: THP-1 cells were treated with QR-si510 siRNA nanoplexes for 12, 48, 96 hr, and up to 1 week and uptake of the nanoplexes was observed using confocal microscopy. (a) Representative local emission spectra confirms that the fluorescence signal in the THP-1 cells is localized to the QR si510 HIV-1siRNA nanoplexes, evident from the sharp emission spectra peak of 625 nm. (b) (A–J): Parallel Fluorescence and transmission images showing fluorescence (red) staining indicating QR-si510 siRNA nanoplex uptake by THP-1 cells.
Mentions: The cellular uptake of the QR-siRNA nanoplexes was studied using confocal microscopy. Figure 6(a) shows the confocal microscopic imaging and the local spectra of THP-1 cells treated with functional QR-si510 HIV-1siRNA nanoplexes. Confocal microscopy images were obtained with laser excitation at 405 nm. Figure 6(a) shows the representative local spectra obtained from different points in the confocal images from cells. The red labeling of THP-1 cells confirms the fluorescence signal is localized to the QR si510 HIV-1siRNA nanoplexes which is evident from the sharp emission spectra peaking around 625 nm. Additionally, it is important to note that the cellular autofluorescence which is normally located around 500 nm shows no interference with the emission from QRs. Our results show uptake of the QR-si510 HIV-1siRNA nanoplexes by the THP-1 cells at 12, 48, 96 hr, and 1 week posttransfection (Figure 6(b) (A)–(J)).

Bottom Line: We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication.Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period.HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

View Article: PubMed Central - PubMed

Affiliation: Division of Allergy, Immunology, and Rheumatology, Department of Medicine, University at Buffalo, The State University of New York, 640 Ellicott Street, Room 444 Innovation Center, Buffalo, NY 14203, USA.

ABSTRACT
HIV-1 replication can be efficiently inhibited by intracellular expression of an siRNA targeting the viral RNA. We used a well-validated siRNA (si510) which targets the poly A/TAR (transactivator of the HIV-1 LTR) site and suppresses viral replication. Nanotechnology holds much potential for impact in the field of HIV-1 therapeutics, and nanoparticles such as quantum rods (QRs) can be easily functionalized to incorporate siRNA forming stable nanoplexes that can be used for gene silencing. We evaluated the efficacy of the QR-si510 HIV-1 siRNA nanoplex in suppressing viral replication in the HIV-1-infected monocytic cell line THP-1 by measuring p24 antigen levels and gene expression levels of HIV-1 LTR. Our results suggest that the QR-si510 HIV-1 siRNA nanoplex is not only effective in delivering siRNA, but also in suppressing HIV-1 viral replication for a longer time period. HIV-1 nanotherapeutics can thus enhance systemic bioavailability and offer multifunctionality.

No MeSH data available.


Related in: MedlinePlus