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Delivery of therapeutic AGT shRNA by PEG-Bu for hypertension therapy.

Wang YQ, Wang F, Deng XQ, Sheng J, Chen SY, Su J - PLoS ONE (2013)

Bottom Line: However, up to today, it is still a major challenge to find safe and efficient non-viral vectors.The structure of PEG-Bu was confirmed by proton nuclear magnetic resonance ((1)H-NMR).Moreover, PEG-Bu could efficiently delivery AGT shRNA to knockdown the AGT expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatrics, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

ABSTRACT
Gene silencing by RNA interference (RNAi) is a promising approach for gene therapy. However, up to today, it is still a major challenge to find safe and efficient non-viral vectors. Previously, we reported PEI-Bu, a small molecular weight PEI derivative, as an efficient non-viral carrier. However, like many PEI-based polymers, PEI-Bu was too toxic. In order to reduce cytotoxicity while maintain or even enhance transfecion efficiency, we modified PEI-Bu with poly(ethylene glycol) (PEG) to obtain PEG-Bu, and used it to delivery a theraputic short hairpin RNA (shRNA) targeting angiotensinogen (AGT) to normal rat liver cells (BRL-3A), which was a key target for the treatment of hypertension. The structure of PEG-Bu was confirmed by proton nuclear magnetic resonance ((1)H-NMR). Gel permeation chromatography (GPC) showed that the weight average molecular weight (Mw) of PEG-Bu was 5880 Da, with a polydispersity of 1.58. PEG-Bu could condense gene cargo into spherical and uniform nanoparticles with particle size (65-88 nm) and zeta potential (7.3-9.6 mV). Interestingly and importantly, PEG-Bu displayed lower cytotoxicity and enhanced tranfection efficiency than PEI-Bu after PEGylation in both normal cells BRL-3A and tumor cells HeLa. Moreover, PEG-Bu could efficiently delivery AGT shRNA to knockdown the AGT expression. To sum up, PEG-Bu would be a promising non-viral vector for delivering AGT shRNA to BRL-3A cells for hypertension therapy.

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Characterization of PEG-Bu/pDNA complexes.A: Agarose gel electrophoresis of PEG-Bu/pDNA complexes at various w/w ratios. 1: Marker; 2: naked pDNA; 3–10: polymer/pDNA complexes at w/w ratios of 0.1,0.3,0.5,1,3,5,10,20. B: Particle size and zeta potential of PEG-Bu/pDNA complexes at various w/w ratios. C: Representative atomic force microscopic (AFM) image of PEG-Bu/pDNA complexes at a w/w ratio of 5.
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pone-0068651-g003: Characterization of PEG-Bu/pDNA complexes.A: Agarose gel electrophoresis of PEG-Bu/pDNA complexes at various w/w ratios. 1: Marker; 2: naked pDNA; 3–10: polymer/pDNA complexes at w/w ratios of 0.1,0.3,0.5,1,3,5,10,20. B: Particle size and zeta potential of PEG-Bu/pDNA complexes at various w/w ratios. C: Representative atomic force microscopic (AFM) image of PEG-Bu/pDNA complexes at a w/w ratio of 5.

Mentions: Gene cargo should be condensed into stable nanoparticle for efficient delivery. Agarose gel electrophoresis was performed to measure the pDNA condensation ability of PEG-Bu. Naked pDNA was used as the control. As displayed in Figure 3A, PEG-Bu could completely retard the migration of pDNA when the w/w ratio was 3, indicating that PEG-Bu/pDNA complexes were completely formed at this w/w ratio. This phenomenon could be explained that the positive charges of PEG-Bu were able to neutralize the negative charges of the phosphate groups in pDNA, thus retarding the pDNA migration. The formation of polymer/pDNA was a necessary step in gene delivery, and this could protect the gene cargo from enzymatic degradation [15], thus prolonging the half-life of gene cargo.


Delivery of therapeutic AGT shRNA by PEG-Bu for hypertension therapy.

Wang YQ, Wang F, Deng XQ, Sheng J, Chen SY, Su J - PLoS ONE (2013)

Characterization of PEG-Bu/pDNA complexes.A: Agarose gel electrophoresis of PEG-Bu/pDNA complexes at various w/w ratios. 1: Marker; 2: naked pDNA; 3–10: polymer/pDNA complexes at w/w ratios of 0.1,0.3,0.5,1,3,5,10,20. B: Particle size and zeta potential of PEG-Bu/pDNA complexes at various w/w ratios. C: Representative atomic force microscopic (AFM) image of PEG-Bu/pDNA complexes at a w/w ratio of 5.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3716693&req=5

pone-0068651-g003: Characterization of PEG-Bu/pDNA complexes.A: Agarose gel electrophoresis of PEG-Bu/pDNA complexes at various w/w ratios. 1: Marker; 2: naked pDNA; 3–10: polymer/pDNA complexes at w/w ratios of 0.1,0.3,0.5,1,3,5,10,20. B: Particle size and zeta potential of PEG-Bu/pDNA complexes at various w/w ratios. C: Representative atomic force microscopic (AFM) image of PEG-Bu/pDNA complexes at a w/w ratio of 5.
Mentions: Gene cargo should be condensed into stable nanoparticle for efficient delivery. Agarose gel electrophoresis was performed to measure the pDNA condensation ability of PEG-Bu. Naked pDNA was used as the control. As displayed in Figure 3A, PEG-Bu could completely retard the migration of pDNA when the w/w ratio was 3, indicating that PEG-Bu/pDNA complexes were completely formed at this w/w ratio. This phenomenon could be explained that the positive charges of PEG-Bu were able to neutralize the negative charges of the phosphate groups in pDNA, thus retarding the pDNA migration. The formation of polymer/pDNA was a necessary step in gene delivery, and this could protect the gene cargo from enzymatic degradation [15], thus prolonging the half-life of gene cargo.

Bottom Line: However, up to today, it is still a major challenge to find safe and efficient non-viral vectors.The structure of PEG-Bu was confirmed by proton nuclear magnetic resonance ((1)H-NMR).Moreover, PEG-Bu could efficiently delivery AGT shRNA to knockdown the AGT expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatrics, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

ABSTRACT
Gene silencing by RNA interference (RNAi) is a promising approach for gene therapy. However, up to today, it is still a major challenge to find safe and efficient non-viral vectors. Previously, we reported PEI-Bu, a small molecular weight PEI derivative, as an efficient non-viral carrier. However, like many PEI-based polymers, PEI-Bu was too toxic. In order to reduce cytotoxicity while maintain or even enhance transfecion efficiency, we modified PEI-Bu with poly(ethylene glycol) (PEG) to obtain PEG-Bu, and used it to delivery a theraputic short hairpin RNA (shRNA) targeting angiotensinogen (AGT) to normal rat liver cells (BRL-3A), which was a key target for the treatment of hypertension. The structure of PEG-Bu was confirmed by proton nuclear magnetic resonance ((1)H-NMR). Gel permeation chromatography (GPC) showed that the weight average molecular weight (Mw) of PEG-Bu was 5880 Da, with a polydispersity of 1.58. PEG-Bu could condense gene cargo into spherical and uniform nanoparticles with particle size (65-88 nm) and zeta potential (7.3-9.6 mV). Interestingly and importantly, PEG-Bu displayed lower cytotoxicity and enhanced tranfection efficiency than PEI-Bu after PEGylation in both normal cells BRL-3A and tumor cells HeLa. Moreover, PEG-Bu could efficiently delivery AGT shRNA to knockdown the AGT expression. To sum up, PEG-Bu would be a promising non-viral vector for delivering AGT shRNA to BRL-3A cells for hypertension therapy.

Show MeSH
Related in: MedlinePlus