EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION
Aim. Evaluation of an antiviral effect of miRNA in the nanoparticles of a polycationic compound against mRNA of vp16 protein (UL48 gene) of herpes simplex virus type 2 (HSV-2) in vitro. Materials and methods. 50% aqueous solution of polyethyleneimine (BDH, Great Britain), chitosan, containing approx...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | Russian |
| Published: |
Central Research Institute for Epidemiology
2015-06-01
|
| Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
| Subjects: | |
| Online Access: | https://microbiol.crie.ru/jour/article/view/14027 |
| _version_ | 1797782183165820928 |
|---|---|
| author | A. S Borisenko O. A Svitich G. G Krivtsov V. F Lavrov V. S Popova T. M Parfenova |
| author_facet | A. S Borisenko O. A Svitich G. G Krivtsov V. F Lavrov V. S Popova T. M Parfenova |
| author_sort | A. S Borisenko |
| collection | DOAJ |
| description | Aim. Evaluation of an antiviral effect of miRNA in the nanoparticles of a polycationic compound against mRNA of vp16 protein (UL48 gene) of herpes simplex virus type 2 (HSV-2) in vitro. Materials and methods. 50% aqueous solution of polyethyleneimine (BDH, Great Britain), chitosan, containing approximately 15% of N-acetylated glucosamine chains (Sonat, Russia), hydrazine-hydrate and other chemical reagents (Chimmed, Russia); Vero continuous cell line, MS HSV-2 virus were used. Vero cells were cultivated in DMEM medium supplemented by 10% fetal bovine serum at 37°C in the atmosphere of 5% CO2. Cell viability was evaluated by using Neutral Red vital stain and MTT-test. Primers and probes for RT-PCR were modeled in Vector NTI 8.0 computer program according to the mRNA sequences of the studied genes (the sequences were obtained from GenBank) and synthesized in Sintol (Russia). RT-PCR tests were set using a standard procedure. Synthesis of PEI-PG-chitosan was carried out by Krivtsov G.G. et al. (2010). Results. A design and synthesis of nucleotide sequences, that have interfering activity against this virus, was carried out to study the effect of siRNA on HSV-2 virus replication. During simultaneous addition of HSV-2 and specific siRNA to Vero cells in cell culture, a significant (by 4 lg) reduction of virus yield was observed. A level of UL48 mRNA expression level was determined after the influence of various siRNA variants. A S2 siRNA variant was shown to cause the most pronounced virus-inhibiting effect, aiming for the center of RNA-target (the level of expression of the studied gene decreased by 0.5 lg). Conclusion. siRNA in the PEI-PG-chitosan complexes were established to possess in vitro pronounced suppressive HSV-2 replication activity. The results obtained could be used in creation of new therapeutic preparation against herpes viruses. |
| first_indexed | 2024-03-13T00:07:24Z |
| format | Article |
| id | doaj.art-112c65db7c8c40d895aee6f16275bab4 |
| institution | Directory Open Access Journal |
| issn | 0372-9311 2686-7613 |
| language | Russian |
| last_indexed | 2024-03-13T00:07:24Z |
| publishDate | 2015-06-01 |
| publisher | Central Research Institute for Epidemiology |
| record_format | Article |
| series | Журнал микробиологии, эпидемиологии и иммунобиологии |
| spelling | doaj.art-112c65db7c8c40d895aee6f16275bab42023-07-12T20:20:08ZrusCentral Research Institute for EpidemiologyЖурнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132015-06-0192331372453EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATIONA. S Borisenko0O. A Svitich1G. G Krivtsov2V. F Lavrov3V. S Popova4T. M Parfenova5Mechnikov Research Institute of Vaccines and SeraMechnikov Research Institute of Vaccines and SeraMechnikov Research Institute of Vaccines and SeraMechnikov Research Institute of Vaccines and SeraMechnikov Research Institute of Vaccines and SeraMechnikov Research Institute of Vaccines and SeraAim. Evaluation of an antiviral effect of miRNA in the nanoparticles of a polycationic compound against mRNA of vp16 protein (UL48 gene) of herpes simplex virus type 2 (HSV-2) in vitro. Materials and methods. 50% aqueous solution of polyethyleneimine (BDH, Great Britain), chitosan, containing approximately 15% of N-acetylated glucosamine chains (Sonat, Russia), hydrazine-hydrate and other chemical reagents (Chimmed, Russia); Vero continuous cell line, MS HSV-2 virus were used. Vero cells were cultivated in DMEM medium supplemented by 10% fetal bovine serum at 37°C in the atmosphere of 5% CO2. Cell viability was evaluated by using Neutral Red vital stain and MTT-test. Primers and probes for RT-PCR were modeled in Vector NTI 8.0 computer program according to the mRNA sequences of the studied genes (the sequences were obtained from GenBank) and synthesized in Sintol (Russia). RT-PCR tests were set using a standard procedure. Synthesis of PEI-PG-chitosan was carried out by Krivtsov G.G. et al. (2010). Results. A design and synthesis of nucleotide sequences, that have interfering activity against this virus, was carried out to study the effect of siRNA on HSV-2 virus replication. During simultaneous addition of HSV-2 and specific siRNA to Vero cells in cell culture, a significant (by 4 lg) reduction of virus yield was observed. A level of UL48 mRNA expression level was determined after the influence of various siRNA variants. A S2 siRNA variant was shown to cause the most pronounced virus-inhibiting effect, aiming for the center of RNA-target (the level of expression of the studied gene decreased by 0.5 lg). Conclusion. siRNA in the PEI-PG-chitosan complexes were established to possess in vitro pronounced suppressive HSV-2 replication activity. The results obtained could be used in creation of new therapeutic preparation against herpes viruses.https://microbiol.crie.ru/jour/article/view/14027small interfering rnaantiviral effectherpes simplex virus type 2polycationic compounds |
| spellingShingle | A. S Borisenko O. A Svitich G. G Krivtsov V. F Lavrov V. S Popova T. M Parfenova EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION Журнал микробиологии, эпидемиологии и иммунобиологии small interfering rna antiviral effect herpes simplex virus type 2 polycationic compounds |
| title | EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION |
| title_full | EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION |
| title_fullStr | EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION |
| title_full_unstemmed | EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION |
| title_short | EFFECTIVENESS OF POLYCATIONIC NANOPARTICLES OF POLYETHYLENEIMINE-POLYHYDROZIDE-CHITOSAN (PEI-PG-OCHG) AS A VECTOR FOR SMALL INTERFERING RNA, DIRECTED TO SUPPRESS HERPES SIMPLEX TYPE 2 VIRUS REPLICATION |
| title_sort | effectiveness of polycationic nanoparticles of polyethyleneimine polyhydrozide chitosan pei pg ochg as a vector for small interfering rna directed to suppress herpes simplex type 2 virus replication |
| topic | small interfering rna antiviral effect herpes simplex virus type 2 polycationic compounds |
| url | https://microbiol.crie.ru/jour/article/view/14027 |
| work_keys_str_mv | AT asborisenko effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication AT oasvitich effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication AT ggkrivtsov effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication AT vflavrov effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication AT vspopova effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication AT tmparfenova effectivenessofpolycationicnanoparticlesofpolyethyleneiminepolyhydrozidechitosanpeipgochgasavectorforsmallinterferingrnadirectedtosuppressherpessimplextype2virusreplication |