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Updated: Oct 16, 2026
A Digital Microfluidic Electroporation Platform for Low-Input CRISPR Genome Editing and mRNA Transfection In Suspension T Cells and 3D Cell Models
Published on: July 17, 2026
Single-Particle Deconvolution of Electroporation Artifacts Guides the Rational Engineering of CD47-EV-Liposome
Xiaozhen Zhan1, Yurou Li1, Chen Chen1
1Department of Chemical Biology, MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, Fujian Key Laboratory of Chemical Biology (Xiamen University), State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian361005, China.
Abstract:
The clinical translation of small interfering RNA (siRNA) therapeutics is hampered by the lack of efficient delivery systems. Extracellular vesicles (EVs) hold great promise as natural carriers, yet conventional loading methods, most notably electroporation, suffer from low encapsulation efficiency, structural damage, and poorly characterized aggregation artifacts. Using single-particle nano-flow cytometry, we deconvoluted these electroporation-induced artifacts and revealed that true loading efficiency is only ∼24%. To overcome these limitations, we engineered CD47-functionalized EV-liposome hybrid vesicles via membrane fusion of CD47-overexpressing EVs with siRNA-loaded liposomes. The hybrid design integrates the high payload capacity of synthetic liposomes with the intrinsic biocompatibility and immune-evasive functionality of CD47-engineered EVs. In a Transwell coculture model simulating the competitive uptake environment of the tumor microenvironment, the hybrid vesicles evaded macrophage clearance and delivered anti-PD-L1 siRNA to tumor cells, resulting in ∼60% PD-L1 gene silencing, three times that of electroporation (∼20%). By providing a rationally designed hybrid nanocarrier with validated immune-evasive and RNAi performance, this work offers a significant advancement for EV-based RNA interference (RNAi) therapeutics.
