PreciScribe T7 RNA Polymerase

Product Descriptions

PreciScribe T7 RNA polymerase is a recombinant T7 RNA polymerase designed through protein engineering, which enhances transcription stability and precision and significantly reduces dsRNA by-products while maintaining other properties of wild-type T7 RNA polymerase including having high specificity for the T7 phage promoters.

Specification & Components

Applications

• Single-stranded RNA synthesis including mRNA, siRNA, sgRNA and other RNA precursors, as well as isotopically labeled or non-isotopically labeled RNA probes.

• Capped mRNA synthesis in reactions containing Cap analogs.

Unit Definition

One Unit of enzyme is defined as the amount of enzyme required to incorporate 1 nmol NTP into RNA within 1 hour at 37℃.

Product Highlights

• PreciScribe T7 RNA Polymerase is an engineered T7 RNA Polymerase, and it maintains high RNA transcription efficiency and yield.

• RNAs produced by PreciScribe T7 RNA Polymerase have very low levels of double-stranded RNA (dsRNA) and less immunogenic.

• Suitable for in vitro RNA synthesis for linear RNA and circular RNA production, and other RNA researches.

Key Performance Data

PreciScribe T7 RNA polymerase significantly reduces dsRNA byproduct

Quantification of dsRNA by-products from different T7 RNA polymerases. Quantification of dsRNA by-products from different T7 RNA polymerase transcripts using J2 as capture antibody and K2 as detection antibody. In (A) blot assay or (B) enzyme-linked immunosorbent assay. PreciScribe T7 RNA polymerase significantly reduces dsRNA byproduct compared to wild-type.

PreciScribe T7 RNA polymerase has a transcriptional yield comparable to wild-type T7 RNA Pol

Quantification of transcription yield of different T7 RNA polymerases. (A) Transcription using 0.5 kb linearized plasmid template at 37 °C for 2 h, with clear bands of single product under 1% agarose gel electrophoresis; (B) PreciScribe T7 RNA polymerase can transcribe RNA at raised temperature (up to 44 C) like wild-type enzyme and with comparable yields.

PreciScribe T7 RNA polymerase has normal co-transcription capping efficiency

Co-transcription capping performance. PreciScribe T7 RNA polymerase co-transcription capping efficiency is the same as that of wild-type T7 RNA Pol.

PreciScribe T7 RNA polymerase is compatible with different modified uridinates

Transcription with modified UTP  PreciScribe T7 RNA polymerase is compatible with different modified uridines and has no effect on (A) incorporation rate and (B) transcription yield.

PreciScribe T7 RNA polymerase is suitable for synthesizing circular RNAs that are functional and have low immunogenicity

Circular RNA synthesis and cell-level expression. (A) PreciScribe T7 RNA polymerase has the same transcriptional yield as wild-type T7 RNA Pol and (B) circular RNA efficiency. (C) The total RNA product of circRNA purified was transfected into cells, and the expression of firefly luciferase (Fluc) was measured. (D) IFN-Beta produced by IVT products obtained by PreciScribe T7 RNA polymerase reaction was significantly lower than that obtained by wild-type T7 RNA Pol reaction.