Semliki Forest Virus (SFV) Self-Amplifying RNA Delivered to J774A.1 Macrophage Lineage by Its Association with a Purified Recombinant SFV Capsid Protein

Author:

Gomes Roselane P.12,Barbosa Flavia F.23,Toledo Marcelo A. S.45,Jorge Soraia A. C.1,Astray Renato M.3ORCID

Affiliation:

1. Viral Biotechnology Laboratory, Butantan Institute, Av. Vital Brasil 1500, São Paulo 05503-900, Brazil

2. Programa Interunidades em Biotecnologia, Universidade de São Paulo, São Paulo 05508-060, Brazil

3. Multipurpose Laboratory, Butantan Institute, Av. Vital Brasil 1500, São Paulo 05503-900, Brazil

4. Department of Hematology, Oncology, Hemostaseology and Stem Cell Transplantation, RWTH Aachen University Medical School, 52074 Aachen, Germany

5. Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf (CIO ABCD), 52074 Aachen, Germany

Abstract

The Semliki Forest virus capsid protein (C) is an RNA binding protein which exhibits both specific and unspecific affinities to single-strand nucleic acids. The putative use of the self-amplifying RNAs (saRNAs) of alphaviruses for biotechnological purpose is one of the main studied strategies concerning RNA-based therapies or immunization. In this work, a recombinant C protein from SFV was expressed and purified from bacteria and used to associate in vitro with a saRNA derived from SFV. Results showed that the purified form of C protein can associate with the saRNA even after high temperature treatment. The C protein was associated with a modified saRNA coding for the green fluorescent protein (GFP) and delivered to murine macrophage cells which expressed the GFP, showing that the saRNA was functional after being associated with the recombinant purified C protein.

Funder

CAPES scholarship

CNPq

FAPESP

Butantan Foundation

Publisher

MDPI AG

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