Controlled Delivery of MicroRNAs into Primary Cells Using Nanostraw Technology

Area of Science:

• Epigenetics
• Molecular Biology
• Biotechnology

Background:

• MicroRNAs (miRNAs) are crucial epigenetic regulators of cellular processes.
• The dosage-dependent effects of miRNAs on gene regulation are largely unexplored.
• Current miRNA delivery methods lack precise dosage control.

Purpose of the Study:

• To develop a delivery platform enabling precise control over intracellular miRNA dosage.
• To investigate the dose-dependent effects of miRNAs on cellular responses.
• To demonstrate efficient miRNA delivery into hard-to-transfect cells.

Main Methods:

• Development of a nanostraw-electroporation platform for miRNA delivery.
• Decoupling miRNA delivery from cellular uptake mechanisms like endocytosis.
• Demonstration of ratiometric intracellular delivery of miR-181a and miR-27a into fibroblasts.
• Assessment of miRNA delivery into primary keratinocytes.

Main Results:

• The nanostraw-electroporation platform precisely controls the amount of miRNA delivered intracellularly.
• Ratiometric delivery of specific miRNAs (miR-181a, miR-27a) into dermal fibroblasts was achieved.
• Efficient miRNA delivery into primary keratinocytes, a challenging cell type, was demonstrated.

Conclusions:

• The nanostraw-electroporation platform overcomes limitations of current miRNA delivery systems.
• This technology enables precise investigation of miRNA dosage-dependent gene regulation.
• It opens new avenues for therapeutic applications involving miRNA modulation, particularly in difficult-to-transfect cells.