2019 / 06

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DOI: 10.24075/brsmu.2019.079

ORIGINAL RESEARCH

Engineering a recombinant Herpesvirus saimiri strain by co-culturing transfected and permissive cells

Hamad A1, Chumakov SP2
About authors

1 Moscow Institute of Physics and Technology, Dolgoprudny, Russia

2 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Russia

Correspondence should be addressed: Stepan P. Chumakov
Miklouho-Maclay, 16/10, Moscow, 117997; moc.liamg@lukhtah

About paper

Funding: this study was supported by the Ministry of Science and Higher Education of the Russian Federation (Project ID RFMEFI60418X0205).

Acknowledgement: we thank to the Center for Precision Genome Editing and Genetic Technologies for Biomedicine (Moscow) for the genetic research methods.

Author contribution: Hamad A — manipulations on cell cultures, molecular cloning; Chumakov SP — study plan; manipulations on viral stocks, transfection, titration, data analysis, manuscript preparation.

Received: 2019-11-11 Accepted: 2019-11-25 Published online: 2019-12-12
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Fig. 1. A schematic representation of the HVS vCD59 region after insertion of the expression cassette. vCD59 flank 1 and vCD59 flank 2 represent recombination homology regions
Fig. 2. A population of permissive OMK cells transduced with the expression cassette SFFV-PURO-OFP
Fig. 3. А. Dynamics of fluorescent populations after HVS-OFP infection. B. A fluorescent plaque formed after OMK cells were infected with the recombinant HVS-OFP. C, D. PBMC (С) and A549 cells (D) transduced with HVS-OFP. E, F. Co-culturing of HEK-293T cells transfected with the recombination cassette and the expressers of the measles virus strain glycoproteins with OMK cells infected with HVS at low (E) and high (F) confluence
Table 1. Primers used for the assembly of the expression cassette
Table 2. The number and ratios of co-cultured cells and the viral titer in the supernatants
Table 3. Conditions for permissive OMK cell line transduction with the expression cassette for HVS recombination and transduction efficacy