T7 RNA POLYMERASE VARIANTS Russian patent published in 2023 - IPC C12N9/12 

FIELD: biotechnology.

SUBSTANCE: invention is a constructed RNA polymerase containing a polypeptide sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with reference sequence SEQ ID NO: 4, where the specified polypeptide sequence contains at least the substitution 664W/F/Y, 404E/Y, 514F/I/L/Y, 661E/Y, 446W/Y, 136E/I, 137W, 357G/K/L/M/N/Q/R/S/T/V/W, 393L/Y, 394R, 397F/M/Q/W/Y, 401V, 444F/H/l/V, 478F/M/W, 513C/F /K/L/R/T/W/Y, 582N, 635F/R/W, 636L, 637G/P/S, 639H, 642L, 643A, 645V, 653C, 656F/W or 660A/C/M/S /T/W/F/N or any combination thereof, and wherein the amino acid positions of the said polypeptide sequence are numbered relative to SEQ ID NO: 4 or 15. The invention also relates to an expression vector and a host cell for producing engineered RNA polymerase.

EFFECT: invention makes it possible to obtain variants of RNA polymerase for selective incorporation of a symmetrically capped GTP analog with respect to GTP during transcription initiation in vitro.

52 cl, 1 dwg, 7 tbl, 7 ex