ISSN 1866-8836
Клеточная терапия и трансплантация

GC-03. Prospects for the use of polyelectrolyte nanocapsules in gene therapy approaches

Tatiana V. Machel1,2,3, Anastasia S. Bukreeva3, Anna S. Rogova3, Ekaterina E. Stefanovskaya3, Oleksii O. Peltek2, Mikhail A. Trofimov4, Yana V. Tarakanchikova3,4, Kirill V. Lepik1, Mikhail V. Zyuzin2, Alexander S. Timin1,3,5, Albert R. Muslimov1,3,4

1 RM Gorbacheva Research Institute of Pediatric Oncology, Hematology and Transplantation, Pavlov University, St. Petersburg, Russia
2 The ITMO University, St. Petersburg, Russia
3 Peter The Great St.Petersburg Polytechnic University, St. Petersburg, Russia
4 St. Petersburg Academic University, St. Petersburg, Russia
5 National Research Tomsk Polytechnic University, Tomsk, Russia

Contact: Dr. Tatiana B. Machel, phone: +7 (999) 212 2336, e-mail:

doi 10.18620/ctt-1866-8836-2020-9-3-1-152



Nowadays, gene therapy is one of the most promising methods of treating infectious, oncological, and hereditary diseases. This method has recently received a new development, due to discovery of genome-editing tools with enormous potential for application because of their high specificity. There are two main approaches to this technology: in vivo, i.e., introduction of gene constructs directly into the patient’s body, and ex vivo treatment by introducing of in vitro transfected donor or patient’s cells. However, the issue of efficient and safe delivery of genetic constructs into relevant cells is a crucial limitation for the widespread introduction of this technology into clinical practice. Viral vectors are already used medically, but they have several limitations, e.g., mutagenesis and inflammatory response and the need to comply with special technical production requirements, which determine high cost of the final product. One of the promising carriers for delivery of safe and efficient biologically active compounds may be polyelectrolyte micro- and nanocapsules obtained by Layer-by-Layer deposition of biodegradable polymer layers of calcium carbonate core with a pre-immobilized delivered component. Compared to others, this method provides many advantages, such as high loading capacity, relatively low manufacturing costs, ease of the particle size regulation, low toxicity, and ability to protect the transferred material from the aggressive effects of biological media in the body. This work was aimed for studying effectiveness of the polyelectrolyte capsules as a platform for genetic material delivery. To this purpose, the following tasks were set and solved:
1. Toxicity testing of the polyelectrolyte capsules;
2. Assessing the packing efficiency of genetic material into the resulting carriers;
3. Modification of polyelectrolyte capsules to increase the efficiency of transfection;
4. Determination of optimal conditions for transfection of primary human macrophages.

Materials and methods

The capsules were prepared by a layering of oppositely charged Polyarginine/Dextran sulfate (PARG/DEXS) and human serum albumin/tannic acid (HSA/TA) polymers, using the Layer-by-Layer technology upon calcium carbonate cores obtained by co-precipitation of sodium carbonate and calcium chloride aqueous solutions. The size and efficiency of genetic material packing into the capsules were assessed using DLS and agarose gel electrophoresis techniques. The following test genetic constructs were used: plasmid DNA and messenger RNA (mRNA) encoding the green fluorescent protein (GFP). HEK293T and THP-1 cell lines, as well as primary human macrophages isolated from blood samples from healthy donors, were used for the in vitro experiments. The efficiency of cell transfection was analyzed by flow cytometry and scanning laser confocal microscopy.


Initially, we have developed a platform for intracellular delivery of genetic material consisting of polyelectrolyte capsules at a size of 500-700 nm. Such carrier particles demonstrated low cytotoxicity (viability of more than 90%) in case of using a 100:1 capsule-to-cell ratio. A high ability of the capsules to retain the delivered material was revealed. Experiments with the plasmid DNA delivery into HEK293T have shown its successful transfection to more than 70% of the cells. Transfection efficiency of mRNAs to the THP-1 cells and primary human macrophages was 60%.


We have demonstrated that polyelectrolyte capsules provide a highly effective and low-toxic platform for in vitro delivery of genetic material to the target cells. Also, this method is easy to use, and it does not require special equipment. In the future, it is planned to conduct experiments for studying potential transfer of clinically relevant genetic constructs into difficult-to-transfect cell lines, and usage of polyelectrolyte capsules for in vivo delivery of genetic material.


The study was carried out with the financial support of the Russian Science Foundation in the framework of a scientific project № 20-45-01012 between Russian Federation and Belgium. A. R. Muslimov thanks the Russian Foundation for Basic Research, grant No.19-015-00098 for the financial support.


Nanocapsules, polyelectrolytes, cell transfection, gene therapy.

Volume 9, Number 3

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doi 10.18620/ctt-1866-8836-2020-9-3-1-152

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