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Related Concept Videos

Experimental RNAi02:15

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

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Related Experiment Video

Updated: May 12, 2026

Molecular Modulation by Lentivirus-Delivered Specific shRNAs in Endoplasmic Reticulum Stressed Neurons
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Molecular Modulation by Lentivirus-Delivered Specific shRNAs in Endoplasmic Reticulum Stressed Neurons

Published on: April 24, 2021

Voltage Preconditioning Allows Modulated Gene Expression in Neurons Using PEI-complexed siRNA.

Arati Sridharan, Chetan Patel, Jit Muthuswamy

    Molecular Therapy. Nucleic Acids
    |March 28, 2013
    PubMed
    Summary

    We developed a voltage-controlled chemical transfection method for high-efficiency siRNA delivery in neurons. This technique enhances polyethylenimine-siRNA complex uptake, improving gene silencing and cell viability for research applications.

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    Subtype-selective Electroporation of Cortical Interneurons
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    Subtype-selective Electroporation of Cortical Interneurons

    Published on: August 18, 2014

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    Published on: April 24, 2021

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    Subtype-selective Electroporation of Cortical Interneurons
    06:42

    Subtype-selective Electroporation of Cortical Interneurons

    Published on: August 18, 2014

    Area of Science:

    • Biotechnology
    • Molecular Biology
    • Neuroscience

    Background:

    • Efficient delivery of small interfering RNA (siRNA) is crucial for gene silencing applications, especially in challenging cell types like neurons.
    • Current transfection methods often suffer from low efficiency, poor viability, or lack of precise control.

    Purpose of the Study:

    • To present a novel voltage-controlled chemical transfection strategy for enhanced siRNA delivery.
    • To optimize polyethylenimine (PEI)-siRNA complex delivery in neuroblastoma (neuro2A) cells and primary neurons.
    • To investigate the underlying mechanism and voltage-dependent modulation of siRNA uptake.

    Main Methods:

    • Applied low voltage pulses to cells prior to administering PEI-siRNA complexes.
    • Quantified transfection efficiency and cell viability using live assays and fluorescently tagged siRNA.
    • Utilized propidium iodide staining to differentiate from electroporation and explored an electro-endocytotic mechanism.
    • Analyzed siRNA uptake and loading via imaging and assessed gene knockdown (GAPDH, BDNF) and cellular changes.

    Main Results:

    • Transfection efficiency in neuro2A cells increased from 62% to 98% at -1V.
    • Transfection efficiency in primary hippocampal neurons increased from 30% to 76% at -1V.
    • siRNA loading showed a fourfold increase at -1V compared to ±2-3V, with negligible transfection at higher voltages.
    • Successful gene knockdown of GAPDH and modulated cell density/dendritic morphology via BDNF knockdown were demonstrated.

    Conclusions:

    • The voltage-controlled chemical transfection method significantly enhances siRNA delivery efficiency and viability in neurons and neuro2A cells.
    • The mechanism appears to be electro-endocytosis, not electroporation, with voltage levels precisely tuning siRNA uptake.
    • This method holds promise for high-throughput screening of siRNA libraries, particularly in difficult-to-transfect neuronal cells.