№ 6 / 2017 / 10-15

STUDY OF RECOMBINANT APOLIPOPROTEIN A-I AS A CARRIER OF PLASMID DNA TO THE RAT HEPATOCYTES

Author: Ryabchenko A.V.¹, Kotova M.V.², Trifonova N.V³, Knyazev R.A.⁴, Polyakov L.M.⁵

Abstract

Purpose of the study was to obtain a modified variant of apolipoprotein A-I and studying its ability to transfer plasmid DNA into rat hepatocytes and comparison with the native form of the protein. Material and methods: gene cloning and production of recombinant proteins in E. coli, incubation of proteins in complex with plasmid pTagGFP2-C in vitro with rat hepatocytes, followed by cell analysis by flow cytometry. Results and discussion. A modified version of apo A-I (10 amino acid residue lysine added at the C-terminus of the protein) penetrated into the hepatocytes, specifically bound to the plasmid DNA, but did not transfect the plasmid into the cells.

Key words

apolipoprotein A-I, transfection, plasmid, hepatocyte

References

Canine B. F., Hatefi A. Development of recombinant cationic polymers for gene therapy research // Adv. Drug Deliv. Rev. 2010. 62. (15). 1524-1529.

Canine B. F., Wang Y., Hatefi A. Biosynthesis and characterization of a novel genetically engineered polymer for targeted gene transfer to cancer cells // J. Control. Release. 2009. 138. (3). 188-196.

Farve G., Tazi K., Le G., Bennis F, Hachem H, Soula G. High density lipoprotein3 bindings sites are related to DNA biosynthesis in the adenocarcinoma cell line A549 // J. Lipid. Res. 1993. 34. (7). 1093-1106.

Jung H.J., Hwang D.S., Wei Q.D., Cha H.J. Carassius auratus-originated recombinant histone H1 C-terminal peptide as gene delivery material // Biotechnol. Prog. // 2008. 24. (1). 17-22.

Nouri F., Wang X., Dorrani M., Karjoo Z., Hatefi A. A recombinant biopolymeric platform for reliable evaluation of the activity of pH-responsive amphiphile fusogenic peptides // Biomacromolecules. 2013. 14. (6). 2033-2040.

Panin L.E., Gimautdinova O.I., Kuznetsov P.A., Velichko E.Yu., Bazaluk V.V. Structure of eukaryotic DNA binding sites for steroid hormone - apolipoprotein A-I complexes // Mol. Biol. (Mosc.) 2007. 41. (4). 583-588.

Panin L.E., Knyazev R.A., Sumenkova D.V., Gushcha R.S., Polyakov L.M. Role of apolipoprotein Е in regulation of protein and nucleic acids biosynthesis in rat hepatocytes culture // Sibirskiy nauchnyy medtsinskiy zhurnal = Siberian Scientific Medical Journal. 2007. 27. (1). 63–66. [In Russian].

Ray M., Tang R., Jiang Z., Rotello V.M. Quantitative tracking of protein trafficking to the nucleus using cytosolic protein delivery by nanoparticle-stabilized nanocapsules // Bioconjug. Chem. 2015. 26. (6). 1004-1007.

Rosenkranz A.A., Ulasov A.V., Slastnikova T.A., Khramtsov Y.V., Sobolev A.S. Use of intracellular transport processes for targeted drug delivery into a specified cellular compartment // Biochemistry (Mosc.). 2014. 79. (9). 928-946.

Ryabchenko A.V., Karavaev V.S., Beklemishev A.B. Comparative structural and immunochemical analysis of recombinant OspC antigens of Novosibirsk Borrelia garinii and Borrelia afzelii isolates // Sibirskiy nauchnyy medtsinskiy zhurnal = Siberian Scientific Medical Journal. 2010. 30. (2). 6–12. [In Russian].

Ryabchenko A.V., Kotova M.V., Polyakov L.M. Designing superproducer of recombinant human apolipoprotein A-I based on Escherichia coli cells // Sibirskiy nauchnyy medtsinskiy zhurnal = Siberian Scientific Medical Journal. 2015. 35. (6). 107–113. [In Russian].

Ryabchenko A.V., Kotova M.V., Tverdohleb N.V., Knyazev R.A., Polyakov L.M. Production and analysis of biological properties of recombinant human apolipoprotein A-I // Bull. Exp. Biol. Med. 2015. 160. (1). 129-133.

Solovieva V.V., Kudryashova N.V., Rizvanov A.A. Transfer of recombinant nucleic acids into cells (transfection) by means of histones and other nuclear proteins // Geny i kletki = Genes and cells. 2011. 6. (3). 29-40. [In Russian].

Soltani F. Sankian, M., Hatefi A., Ramezani M. Development of a novel histone H1-based recombinant fusion peptide for targeted non-viral gene delivery // Int. J. Prarm. 2013. 441 (1). 307-315.

About Authors (Correspondence):

Polyakov L.M. – doctor of medical sciences, professor, head of the laboratory of medical biotechnology, e-mail: plm@niibch.ru

Full Text

snmzh_6-2017_ryabchenko_i_dr.pdf

Received: 10/01/2018