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 6 publications

(1 citation statement)

References 35 publications

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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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Genetically Encoded Trensor Circuits Report HeLa Cell Treatment with Polyplexed Plasmid DNA and Small-Molecule Transfection Modulators

Redwood-Sawyerr,

Howe,

Evans Theodore

et al. 2024

ACS Synth. Biol.

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HeLa cell transfection with plasmid DNA (pDNA) is widely used to materialize biologicals and as a preclinical test of nucleic acid-based vaccine efficacy. We sought to genetically encode mammalian transfection sensor (Trensor) circuits and test their utility in HeLa cells for detecting molecules and methods for their propensity to influence transfection. We intended these Trensor circuits to be triggered if their host cell was treated with polyplexed pDNA or certain small-molecule modulators of transfection. We prioritized three promoters, implicated by others in feedback responses as cells import and process foreign material and stably integrated each into the genomes of three different cell lines, each upstream of a green fluorescent protein (GFP) open reading frame within a transgene. All three Trensor circuits showed an increase in their GFP expression when their host HeLa cells were incubated with pDNA and the degraded polyamidoamine dendrimer reagent, SuperFect. We next experimentally demonstrated the modulation of PEI-mediated HeLa cell transient transfection by four different small molecules, with Trichostatin A (TSA) showing the greatest propensity to boost transgene expression. The Trensor circuit based on the TRA2B promoter (Trensor-T) was triggered by incubation with TSA alone and not the other three small molecules. These data suggest that mammalian reporter circuits could enable low-cost, high-throughput screening to identify novel transfection methods and reagents without the need to perform actual transfections requiring costly plasmids or expensive fluorescent labels.

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

Cited by 6 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)

Genetically Encoded Trensor Circuits Report HeLa Cell Treatment with Polyplexed Plasmid DNA and Small-Molecule Transfection Modulators

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

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