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Updated: Jun 11, 2025
Lentiviral Mediated Delivery of shRNAs to hESCs and NPCs Using Low-cost Cationic Polymer Polyethylenimine PEI
Published on: May 24, 2022
Plate centrifugation enhances the efficiency of polyethylenimine-based transfection and lentiviral infection
Shaozhe Yang1, Qingwei Zhang1, Yuan Zhuang1
1Henan Key Laboratory of Fertility Protection and Aristogenesis, Luohe Central Hospital, 54 Renmin East Road, Luohe 462000, PR China; Prenatal Diagnostic Center, Luohe Central Hospital, 54 Renmin East Road, Luohe 462000, PR China.
Purpose:
To propose an efficient, reproducible, and consistent transgenic technology based on plate centrifugation, which is particularly useful for polyethylenimine (PEI) transfection and lentiviral infection.
Methods:
We optimized multiple factors that could contribute to transfection efficiency, such as the dosage of the PEI or DNA, the working solution buffer used for diluting the PEI or DNA, the incubation time for the PEI/DNA complexes, and the transfection time.
Results:
Plate centrifugation led to a 5.46-fold increase in the transfection efficiency of PEI-based transfection while maintaining the cell survival rate. Moreover, the average copy number of viral genes in each genome increased 4.96-fold with plate centrifugation. Plate centrifugation alters the spatial arrangement of the PEI/DNA complexes or lentiviruses, increasing the chances of these complexes or viruses coming into contact with target cells, ultimately resulting in improved transfection or infection efficiency.
Conclusions:
We present a protocol based on plate centrifugation for transfection or lentiviral infection that is suitable for genetic modification of primary cells or stem cells.
