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Highly Efficient Messenger RNA Transfection of Hard-to-Transfect Cells using Carbon Nanodots.

ABSTRACT: Although messenger RNA (mRNA)-based therapeutics opened up new avenues for treating various diseases, intracellular delivery of mRNA is still challenging, especially to hard-to-transfect cells. For successful mRNA therapy, the development of a delivery vehicle that can effectively transport mRNA into cells is essential. In this study, we synthesized carbon nanodots (CNDs) as an efficient mRNA delivery vehicle via a one-step microwave-assisted method. CNDs easily formed complexes with mRNA molecules by electrostatic interactions, and the gene delivery performance of CNDs was highly effective in hard-to-transfect cells. Considering their outstanding transfection ability, CNDs are expected to be further applied for mRNA-based cellular engineering.

SUBMITTER: Shin H

PROVIDER: S-EPMC10433478 | biostudies-literature | 2023 Aug

REPOSITORIES: biostudies-literature

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Highly Efficient Messenger RNA Transfection of Hard-to-Transfect Cells using Carbon Nanodots.

Shin Hojeong H Min Dal-Hee DH

ACS omega 20230802 32

Although messenger RNA (mRNA)-based therapeutics opened up new avenues for treating various diseases, intracellular delivery of mRNA is still challenging, especially to hard-to-transfect cells. For successful mRNA therapy, the development of a delivery vehicle that can effectively transport mRNA into cells is essential. In this study, we synthesized carbon nanodots (CNDs) as an efficient mRNA delivery vehicle via a one-step microwave-assisted method. CNDs easily formed complexes with mRNA molecu ...[more]

PMID: 37599943

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Project description:BackgroundThe discovery and development of RNA interference has made a tremendous contribution to the biochemical and biomedical field. However, liposomal transfection protocols to deliver siRNAs to certain types of cells, eg, immune cells, are not viable due to exceedingly low transfection efficiency. While viral delivery and electroporation are two widely adopted approaches to transfect immune cells, they are associated with certain drawbacks such as complexity of preparation, biosafety issues, and high cytotoxicity. We believe amendments can be made to liposomal formulas and protocols to achieve a highly efficient knockdown of genes by liposome-loaded siRNAs.AimThe aim of this study was to use the apoptotic-mimic Ca-PS lipopolyplex to achieve highly efficient siRNA knockdown of genes in the hard-to-transfect macrophages with reduced cytotoxicity and more efficient cellular uptake.ResultsWe devised an anionic liposomal formula containing phosphatidylserine to mimic the apoptotic body, the Ca-PS lipopolyplex. Ca-PS lipopolyplex was proven to be capable of delivering and effecting efficient gene knockdown in multiple cell lines at lowered cytotoxicity. Among the two types of macrophages, namely Ana-1 and bone-marrow derived macrophages, Ca-PS lipopolyplex showed an improvement in knockdown efficiency, as high as 157%, over Lipo2000. Further investigations revealed that Ca-PS promotes increased cellular uptake, lysosomal escape and localization of siRNAs to the perinuclear regions in macrophages. Lastly, transfection by Ca-PS lipopolyplex did not induce spontaneous polarization of macrophages.ConclusionThe apoptotic body-mimic Ca-PS lipopolyplex is a stable, non-cytotoxic liposomal delivery system for siRNAs featuring vastly improved potency for macrophages and lowered cytotoxicity. It is speculated that Ca-PS lipopolyplex can be applied to other immune cells such as T cells and DC cells, but further research efforts are required to explore its promising potentials.

Dataset Information

Highly Efficient Messenger RNA Transfection of Hard-to-Transfect Cells using Carbon Nanodots.

Publications

Highly Efficient Messenger RNA Transfection of Hard-to-Transfect Cells using Carbon Nanodots.

Similar Datasets

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