BIONTECH RNA PHARMACEUTICALS GMBH;TRON - TRANSLATIONALE ONKOLOGIE AN DER UNIVERSITÄTSMEDIZIN DER JOHANNES GUTENBERG-UNIVERSITÄT MAINZ GEMEINNÜTZIGE GMBH
发明人:
BEISSERT, Tim,SAHIN, Ugur,PERKOVIC, Mario
申请号：
EPEP2017/055813
公开号：
WO2017/162461A1
申请日:
2017.03.13
申请国别(地区):
EP
年份:
2017
代理人:
摘要：
The present invention generally relates to systems and methods suitable for high-level protein production. While one or more elements of the present invention are derived from an alphavirus, the present invention does not require propagation of virus particles. In particular, a system comprising two separate RNA molecules is foreseen, each comprising a nucleotide sequence derived from an alphavirus: one RNA molecule comprises a RNA construct for expressing alphavirus replicase, and one RNA molecule comprises a RNA re pi icon that can be replicated by the replicase in trans. The RNA construct for expressing alphavirus replicase comprises a 5'-cap. It was surprisingly found that the 5'-cap is suitable for efficiently driving expression of a transgene from the replicon in trans. The system of the present invention enables expression of a protein of interest in a cell or organism, but is not associated with undesired virus-particle formation. Therefore, the present invention is suitable for efficiently and safely producing a protein of interest, e.g. a therapeutic protein or an antigenic protein, such as a vaccine, in a target organism. Respective methods of protein production in vitro and in vivo as well as medical uses are provided herein. The present invention also provides DNA encoding the RNA molecules of the invention, and cells comprising the RNA molecules of the invention.La présente invention concerne de manière générale des systèmes et des procédés appropriés pour une production protéinique de niveau élevé. Tandis qu'un ou plusieurs éléments de la présente invention sont dérivés d'un alphavirus, la présente invention ne nécessite pas de propagation de particules virales. En particulier, un système comprenant deux molécules d'ARN séparées est prévu, comprenant chacune une séquence nucléotidique dérivée d'un alphavirus : une molécule d'ARN comprend une construction d'ARN pour exprimer une réplicase d'alphavirus et une molécule d'ARN comprend un réplicon d'ARN qu
