Viral Biotechnology Laboratory, Butantan Institute, Av. Vital Brasil 1500, São Paulo 05503-900, Brazil.
Programa Interunidades em Biotecnologia, Universidade de São Paulo, São Paulo 05508-060, Brazil.
Int J Mol Sci. 2024 Jul 18;25(14):7859. doi: 10.3390/ijms25147859.
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.
塞姆利基森林病毒衣壳蛋白(C)是一种 RNA 结合蛋白,对单链核酸表现出特异性和非特异性亲和力。利用甲病毒的自我扩增 RNA(saRNA)用于生物技术目的是 RNA 治疗或免疫的主要研究策略之一。在这项工作中,SFV 的重组 C 蛋白从细菌中表达和纯化,并用于体外与 SFV 衍生的 saRNA 结合。结果表明,即使经过高温处理,纯化的 C 蛋白形式也可以与 saRNA 结合。C 蛋白与编码绿色荧光蛋白(GFP)的修饰 saRNA 结合,并递送至表达 GFP 的鼠巨噬细胞,表明与重组纯化 C 蛋白结合后 saRNA 具有功能。