An efficient method for in vitro gene delivery via regulation of cellular endocytosis pathway

Luo, Jing; Li, Caixia; Chen, Jianlin; Wang, Gang; Gao, Rong; Gu, Zhongwei

doi:10.2147/ijn.s77527

Cited by 5 publications

(1 citation statement)

References 35 publications

(37 reference statements)

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“…These diverse applications of PEI are a consequence of its chemical structure. Due to its numerous positive charges, it can easily enter the cell membrane via endocytosis, which makes it an optimal agent for gene transfer [4]. PEI is a polycationic polymer having linear and branched forms with different molecular weights [5].…”

Section: Introductionmentioning

confidence: 99%

Study of the Intrinsic Fluorescence of a Highly Branched Cationic Dendrimer, Poly(Ethyleneimine) (PEI)

et al. 2019

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Poly(ethyleneimine) (PEI) is a weakly basic, synthetic, polycationic polymer, due to the presence of primary, secondary, and tertiary amino groups. The amino groups are responsible for the variety of applications of PEI (e.g., transfection, bioimaging, solar cell, etc.). Our study presents some new and reproducible methods for the quantification of molecular or mass concentration of highly branched PEI of different molecular weights (800–2000–25,000–750,000 MW PEI). In the course of the direct method, spectrophotometry and fluorometry were applied to determine the absorption and fluorescence of PEI dilution series. An increase in the MW at the same concentration produces a higher count number because of the higher number of amino groups in PEI molecules. The character of increment in fluorescence intensity is essentially different in the case of mass concentrations and molar concentrations. The increment of the fluorescence intensity related to the molar concentration is non-linear. In the case of mass concentration, the slope is linear. Moreover, their fluorescence is enhanced with the decrease in pH values. The spectrophotometry is a reliable method for measuring the quantity of PEI molecules in solution. Our data help in recognizing the detailed properties of PEI in dendrimer research.

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Section: Introductionmentioning

confidence: 99%

Study of the Intrinsic Fluorescence of a Highly Branched Cationic Dendrimer, Poly(Ethyleneimine) (PEI)

et al. 2019

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Genetic variation of the HIV-1 subtype C transmitted/founder viruses long terminal repeat elements and the impact on transcription activation potential and clinical disease outcomes

et al. 2023

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A genetic bottleneck is a hallmark of HIV-1 transmission such that only very few viral strains, termed transmitted/founder (T/F) variants establish infection in a newly infected host. Phenotypic characteristics of these variants may determine the subsequent course of disease. The HIV-1 5’ long terminal repeat (LTR) promoter drives viral gene transcription and is genetically identical to the 3’ LTR. We hypothesized that HIV-1 subtype C (HIV-1C) T/F virus LTR genetic variation is a determinant of transcriptional activation potential and clinical disease outcome. The 3’LTR was amplified from plasma samples of 41 study participants acutely infected with HIV-1C (Fiebig stages I and V/VI). Paired longitudinal samples were also available at one year post-infection for 31 of the 41 participants. 3’ LTR amplicons were cloned into a pGL3-basic luciferase expression vector, and transfected alone or together with Transactivator of transcription (tat) into Jurkat cells in the absence or presence of cell activators (TNF-α, PMA, Prostratin and SAHA). Inter-patient T/F LTR sequence diversity was 5.7% (Renge: 2–12) with subsequent intrahost viral evolution observed in 48.4% of the participants analyzed at 12 months post-infection. T/F LTR variants exhibited differential basal transcriptional activity, with significantly higher Tat-mediated transcriptional activity compared to basal (p<0.001). Basal and Tat-mediated T/F LTR transcriptional activity showed significant positive correlation with contemporaneous viral loads and negative correlation with CD4 T cell counts (p<0.05) during acute infection respectively. Furthermore, Tat-mediated T/F LTR transcriptional activity significanly correlated positively with viral load set point and viral load; and negatively with CD4 T cell counts at one year post infection (all p<0.05). Lastly, PMA, Prostratin, TNF-α and SAHA cell stimulation resulted in enhanced yet heterologous transcriptional activation of different T/F LTR variants. Our data suggest that T/F LTR variants may influence viral transcriptional activity, disease outcomes and sensitivity to cell activation, with potential implications for therapeutic interventions.

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<p>Cationic <em>Antheraea pernyi</em> Silk Fibroin-Modified Adenovirus-Mediated ING4 and IL-24 Dual Gene Coexpression Vector Suppresses the Growth of Hepatoma Carcinoma Cells</p>

Qu¹,

Wang²,

Feng³

et al. 2019

IJN

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IntroductionCancer gene therapy requires both effective tumor suppressor genes and safe vectors that express target genes efficiently. Inhibitor of growth 4 (ING4) inhibits tumor growth via multiple pathways. Interleukin-24 (IL-24) also has tumor-suppressive activity against a broad spectrum of human cancers. Adenovirus (Ad) vectors exhibit high infection efficiency, but potential toxicity related to high doses of adenovirus has led to careful reconsideration of their use in human clinical trials. Antheraea pernyi silk fibroin (ASF) is a cytocompatible and biodegradable natural polymer, and it possesses Arg–Gly–Asp sequences exhibiting a high binding affinity and selectivity for αvβ3 and αvβ5 integrin receptors, which are overexpressed in tumor vessels and most tumor cells.MethodsIn this study, an Arg-Gly-Asp peptide-modified Ad vector coexpressing ING4 and IL-24 was constructed by homologous recombination of the dual gene coexpression transfer plasmid and RGD-modified pAdEasy-1 adenoviral backbone plasmid. The cationic ASF (CASF) was prepared by modifying ASF with low-molecular-weight PEI. The negatively charged Ad vector was modified with CASF to form a CASF/Ad complex.ResultsHuman hepatoma carcinoma SMMC-7721 cells and normal hepatic L-02 cells were infected with the CASF/Ad complex, which showed significantly higher infection efficiency than the naked Ad. The CASF/Ad complex could effectively mediate the expression of the target gene ING4 in SMMC-7721 cells and the secretion of the target gene IL-24 from SMMC-7721 cells, thus inducing apoptosis of hepatoma carcinoma SMMC-7721 cells. The viability of SMMC-7721 and L-02 cells infected with the CASF/Ad complex was further assessed, and it was found that the growth of SMMC-7721 cells was significantly inhibited but that the growth and proliferation of L-02 cells were not affected.ConclusionThe CASF/Ad complex constructed in this study, showing improved infection efficiency and enhanced suppressive effects on human hepatoma carcinoma SMMC-7721 cells, has the potential to reduce the dose of adenovirus and still maintain high infection efficiency and tumor inhibition.

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An efficient method for in vitro gene delivery via regulation of cellular endocytosis pathway

Cited by 5 publications

References 35 publications

Study of the Intrinsic Fluorescence of a Highly Branched Cationic Dendrimer, Poly(Ethyleneimine) (PEI)

Study of the Intrinsic Fluorescence of a Highly Branched Cationic Dendrimer, Poly(Ethyleneimine) (PEI)

Genetic variation of the HIV-1 subtype C transmitted/founder viruses long terminal repeat elements and the impact on transcription activation potential and clinical disease outcomes

<p>Cationic <em>Antheraea pernyi</em> Silk Fibroin-Modified Adenovirus-Mediated ING4 and IL-24 Dual Gene Coexpression Vector Suppresses the Growth of Hepatoma Carcinoma Cells</p>

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