公开(公告)号：US20240060107A1

公开(公告)日：2024-02-22

申请号：US18069123

申请日：2022-12-20

申请人： GENSCRIPT USA INC.

发明人： Cheng-Hsien WU ,  Fengmei PI ,  Robert DEMPCY ,  Shambhavi SHUBHAM ,  Kristine BIELECKI ,  Aaron BALL

IPC分类号： C12P19/34 ,  C12N9/12 ,  C12N9/64 ,  C12N9/16

CPC分类号： C12P19/34 ,  C12N9/1276 ,  C12Y207/07049 ,  C12N9/6472 ,  C12Y304/22056 ,  C12N9/16 ,  C12Y301/03002

摘要： A method for in vitro transcription of a DNA template into RNA includes providing a mixture containing a buffer substance, ribonucleoside triphosphates (NTPs), one or more magnesium salts in a concentration of from about 2 mM to about 60 mM, the DNA template, and a recombinant RNA polymerase, and incubating the reaction mixture at from about 25° C. to about 40° C. for from about 1 hour to about 12 hours thereby producing the RNA. A method for in vitro transcription includes providing a DNA template and a cap analogue that binds to −1 and/or +1 nucleotides of promoter for in vitro transcription, thus producing more full length mRNAs, allowing for more flexibility on the choice of first mRNA base, and providing +2 position open for custom sequence.