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Nanochannel Electro-Injection as a Versatile Platform for Efficient RNA/DNA Programming on Dendritic Cells.

Jing Liu1,2, Juan Jiang1, Caiguanxi Deng1

  • 1Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, P. R. China.

Small (Weinheim an Der Bergstrasse, Germany)
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Summary

Nanochannel electro-injection safely and efficiently delivers nucleic acids into dendritic cells (DCs) for cancer vaccines. This method optimizes transfection efficiency and cell viability, paving the way for advanced DC vaccine development.

Keywords:
DNA/RNAdendritic cellsintracellular deliverynanochannel electro-injection

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Area of Science:

  • Biotechnology
  • Immunotherapy
  • Nanotechnology

Background:

  • Dendritic cell (DC) vaccines are crucial for cancer immunotherapy.
  • Efficient and safe delivery of nucleic acids into DCs is vital for DC vaccine development but remains challenging.
  • Current methods often struggle to balance transfection efficiency with biosafety and preventing DC maturation.

Purpose of the Study:

  • To develop and optimize a novel nanochannel electro-injection (NEI) system for efficient and safe nucleic acid delivery into dendritic cells.
  • To evaluate the NEI system's performance in transfecting various nucleic acid types (DNA, mRNA, circRNA) and its impact on DC viability and maturation.
  • To assess the potential of NEI for in vitro transformation of DCs for cancer vaccine applications.

Main Methods:

  • Utilized a nanochannel electro-injection (NEI) system based on track-etched nanochannel membranes.
  • Localized electric fields on the cell membrane using nano-sized channels for low-voltage electroporation (<30 V).
  • Optimized pulse conditions for transfecting DC2.4 cells with fluorescent dyes, plasmid DNA, messenger RNA, and circular RNA (circRNA).

Main Results:

  • Achieved high transfection efficiency (>70%) and biosafety (viability >85%) for various nucleic acids in DC2.4 cells using NEI.
  • Successfully transfected primary mouse bone marrow DCs with circRNA with 68.3% efficiency.
  • Demonstrated that NEI did not significantly affect cellular viability or induce DC maturation in primary cells.

Conclusions:

  • The nanochannel electro-injection (NEI) system provides a safe and efficient platform for in vitro dendritic cell transformation.
  • NEI overcomes key challenges in nucleic acid delivery for DC vaccine development.
  • This technology holds significant promise for advancing the creation of effective dendritic cell-based cancer vaccines.