GMP-Grade T7 RNA Polymerase

T7 RNA Polymerase (T7RNAP) is a protein encoded by phage T7 DNA recombinantly expressed in Escherichia coli. T7RNAP, a DNA-dependent 5'→3' RNA polymerase, highly specifically recognizes the T7 promoter sequence (5'-TAATACGACTCACTATAG-3'). The template is single-stranded or double-stranded DNA containing the T7 promoter sequence. T7RNAP synthesizes an RNA strand using NTPs, complementary to the DNA template, downstream of the promoter.

We engineered our T7 RNA Polymerase using Structure Aided Multiplex Screening (SAMS), our unique innovative functional recombinant protein engineering platform. It has also undergone optimization of the E. coli expression system, purification process, and buffer formulation. We've designed our T7 to produce low error rates, high efficiency, and high processivity. We've optimized the enzyme for yield and purity, making it both practical and affordable for large-scale production of mRNA / mRNA synthesis.

We manufacture our T7 RNA Polymerase in our cGMP compliant facility. Moreover, we've submitted our GMP T7RNAP to the FDA Drug Master Files (DMF #037660). Customizable regulatory support documentation including batch production records, stability testing, CoA, etc. is available.

Customizable GMP Documentation Package:

→ Datasheet
→ CoA
→ CoO
→ MSDS
→ Melamine Statement
→ TSE/BSE Statement
→ Nitrosamine Statement
→ DMF Filing

Figure 1. After synthesizing mRNA in an IVT reaction, we assessed RNA transcript yield using nanodrop and Agilent 4150 tapestation. We assessed purity using capillary electrophoresis (CE). Overall, yield and purity of our T7RNAP was comparable to leading suppliers.

Figure 2. dsRNA measured after in vitro transcription using J2-based ELISA. KACTUS T7 has low dsRNA contamination comparable or superior to leading competitors.

Figure 3. After synthesis of mRNA, in vitro transcription yield of T7 RNA Polymerase measured with Agilent 4150 Tapestation. KACTUS T7 had 65% higher yield than leading supplier.

Figure 4. Detection of T7 RNA Polymerase by molecular beacon. We analyzed three batches of T7 RNA Polymerase and found similar reaction kinetics across all batches.