公开(公告)号：US20220396798A1

公开(公告)日：2022-12-15

申请号：US17489357

申请日：2021-09-29

申请人： Shi-Lung LIN ,  Samantha CHANG-LIN ,  Jack SK CHEN ,  David TS WU ,  Chia-Ning SHEN ,  Mei-Jung WANG

发明人： Shi-Lung LIN ,  Samantha CHANG-LIN ,  Jack SK CHEN ,  David TS WU ,  Chia-Ning SHEN ,  Mei-Jung WANG

IPC分类号： C12N15/113 ,  C12N15/85

摘要： This invention relates to a novel mRNA composition and its production method useful for developing and manufacturing RNA-based anti-viral and/or anti-cancer vaccines and medicines. This invention includes two types of mRNA constructs, namely “5′-hairpin messenger RNA (5hmRNA)” and “messenger-hairpin-messenger RNA (mhmRNA)”, respectively. Both of 5hmRNA and mhmRNA contain at least a hairpin-like stem-loop RNA structure. The 5hmRNA contains at least a stem-loop RNA structure in the 5′-UTR of a protein/peptide-coding mRNA, while the mhmRNA contains a middle stem-loop structure flanked with two protein/peptide-coding mRNA sequences on both sides. In mhmRNA, the first 5′-mRNA preferably encodes an RNA replicase, for amplifying the second 3′-mRNA in transfected cells. After transfection into target cells, 5hmRNA and mhmRNA can be further translated into at least a desired protein/peptide. To produce highly structured 5hmRNA and mhmRNA, a novel PCR-IVT methodology has been developed and used with a specially designed RNA polymerase-helicase mixture reaction