Skip to Content
MilliporeSigma
  • Enhanced shRNA delivery and ABCG2 silencing by charge-reversible layered nanocarriers.

Enhanced shRNA delivery and ABCG2 silencing by charge-reversible layered nanocarriers.

Small (Weinheim an der Bergstrasse, Germany) (2014-10-22)
Zhenzhen Chen, Lifen Zhang, Yuling He, Youqing Shen, Yanfeng Li
ABSTRACT

Polycationic vectors have been used to deliver short hairpin RNAs (shRNAs) to knock-down genes for cancer therapies, but their inefficiency in lysosomal escape and shRNA release causes their low gene transcription efficiency. Herein, a three-layered polyethyleneimine (PEI)-coated gold nanocomplex interlaid with a pH-responsive charge-reversible chitosan-aconitic anhydride (CS-Aco) is constructed: a Au-PEI/CS-Aco/PEI/shRNA nanoparticle. The negatively charged CS-Aco hydrolyzes into positively charged CS in lysosomes, causing the nanocomposite to disassemble. The released Au-PEI nanoparticles efficiently rupture the lysosomes and thus release the PEI/shRNA polyplexes into cytoplasm, where they quickly disassociate because the PEI chains are short (1.2 kDa). As a consequence, the nanocomplexes display higher shRNA delivery efficiency than the 25 kDa PEI, and efficiently deliver shABCG2 to tumors and markedly silence ABCG2 expression, which sensitizes HepG2 cells to the drugs with minimal toxicity.

MATERIALS
Product Number
Brand
Product Description

Supelco
HEPES, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
SAFC
HEPES
SAFC
HEPES
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 500 μg/mL in H2O
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 10 mg/mL in H2O
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, 98%
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Sigma-Aldrich
HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC)
Sigma-Aldrich
HEPES, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
Ethidium bromide solution, for fluorescence, ~1% in H2O
Sigma-Aldrich
Tricarballylic acid, 99%
Sigma-Aldrich
Ethidium bromide solution, 10 mg/mL
Sigma-Aldrich
Ethidium bromide, ≥95.0%
Sigma-Aldrich
HEPES buffer solution, 1 M in H2O
Sigma-Aldrich
cis-Aconitic anhydride, 95%
Sigma-Aldrich
Ethidium bromide, ~95% (HPLC)
Sigma-Aldrich
Ethidium bromide, BioReagent, for molecular biology, powder