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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

The time-dependent cytotoxicity of QR-si510 siRNA nanoplex on THP-1 cells using the MTT assay. Our results show absence of any toxicity in THP-1 monocytic cultures treated with lipofectamine transfected scrambled siRNA, Lipofectamine transfected si510 HIV-1 siRNA and  the QR-si510 siRNA nanoplex as compared to the untransfected control over a 12 hr to 1 week time period. The concentration of both the scrambled siRNA and the si510 HIV-1 siRNA were 10 nM.  Also included in the MTT assay were THP-1 cells not treated with the HIV-1 virus (no virus control). The results shown are mean ± SD of 3 separate experiments.
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fig2: The time-dependent cytotoxicity of QR-si510 siRNA nanoplex on THP-1 cells using the MTT assay. Our results show absence of any toxicity in THP-1 monocytic cultures treated with lipofectamine transfected scrambled siRNA, Lipofectamine transfected si510 HIV-1 siRNA and the QR-si510 siRNA nanoplex as compared to the untransfected control over a 12 hr to 1 week time period. The concentration of both the scrambled siRNA and the si510 HIV-1 siRNA were 10 nM. Also included in the MTT assay were THP-1 cells not treated with the HIV-1 virus (no virus control). The results shown are mean ± SD of 3 separate experiments.

Mentions: An MTT assay was done to evaluate the effect of the nanoplexes on cell viability of the THP-1 cells. Cell viability was also tested in the THP-1 cells prior to being infected by HIV-1 as well as after 7 days post infection and over 99% viability was observed in cells prior to be treated with the nanoplexes. THP-1 cell viability was measured after treatment with the nanoplexes at 12, 24, 96 hr, and upto 1 week posttransfection using the maximum dose (100 nM) of QR-si510 HIV-1 siRNA. Our results (Figure 2) show greater than 90% viability in THP-1 cells treated with QR-si510 HIV-1 siRNA nanoplexes and under all treatment conditions for up to 1 week. These results demonstrate that the QR-mediated delivery of siRNA did not produce any appreciable cytotoxicity in THP-1 cells.


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)

The time-dependent cytotoxicity of QR-si510 siRNA nanoplex on THP-1 cells using the MTT assay. Our results show absence of any toxicity in THP-1 monocytic cultures treated with lipofectamine transfected scrambled siRNA, Lipofectamine transfected si510 HIV-1 siRNA and  the QR-si510 siRNA nanoplex as compared to the untransfected control over a 12 hr to 1 week time period. The concentration of both the scrambled siRNA and the si510 HIV-1 siRNA were 10 nM.  Also included in the MTT assay were THP-1 cells not treated with the HIV-1 virus (no virus control). The results shown are mean ± SD of 3 separate experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: The time-dependent cytotoxicity of QR-si510 siRNA nanoplex on THP-1 cells using the MTT assay. Our results show absence of any toxicity in THP-1 monocytic cultures treated with lipofectamine transfected scrambled siRNA, Lipofectamine transfected si510 HIV-1 siRNA and the QR-si510 siRNA nanoplex as compared to the untransfected control over a 12 hr to 1 week time period. The concentration of both the scrambled siRNA and the si510 HIV-1 siRNA were 10 nM. Also included in the MTT assay were THP-1 cells not treated with the HIV-1 virus (no virus control). The results shown are mean ± SD of 3 separate experiments.
Mentions: An MTT assay was done to evaluate the effect of the nanoplexes on cell viability of the THP-1 cells. Cell viability was also tested in the THP-1 cells prior to being infected by HIV-1 as well as after 7 days post infection and over 99% viability was observed in cells prior to be treated with the nanoplexes. THP-1 cell viability was measured after treatment with the nanoplexes at 12, 24, 96 hr, and upto 1 week posttransfection using the maximum dose (100 nM) of QR-si510 HIV-1 siRNA. Our results (Figure 2) show greater than 90% viability in THP-1 cells treated with QR-si510 HIV-1 siRNA nanoplexes and under all treatment conditions for up to 1 week. These results demonstrate that the QR-mediated delivery of siRNA did not produce any appreciable cytotoxicity in THP-1 cells.

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