Efficiently Reduce Time, Ensure Quality with the Latest Upgrade to Onepot mRNA Transcription Process !

In recent years, the development and application of mRNA technology have continued to attract attention. mRNA drugs involve inoculating mRNA that encodes antigen proteins into the human body. By utilizing the genetic material inside human cells, they express and synthesize antigen proteins. This process induces and activates the body's immune system through antigen proteins, aiming to prevent and treat diseases. mRNA drugs have advantages such as safety, efficiency, and a short cycle. They can simultaneously induce humoral and cellular immunity and have been applied in various indications, including tumors, infectious diseases, rare diseases, cardiovascular diseases, and more.

The entire process of mRNA drug development can be roughly divided into several steps:

Sequence determination & design—mRNA in vitro transcription—encapsulation preparation

The conventional steps of mRNA in vitro transcription (IVT) include:

Plasmid DNA extraction & linearizationPlasmid DNA purification—In vitro transcription (co-transcriptional capping)—Purification—mRNA stock solution

The onepot mRNA in vitro transcription (IVT) process include:

Plasmid DNA extraction & linearizationIn vitro transcription (co-transcriptional capping)—Purification—mRNA stock solution

Currently, most existing processes involve a single purification step for plasmid DNA after enzymatic cleavage before the IVT step. Yeasen Biotechlogy, based on a mature mRNA application development center, has developed the "Onepot mRNA Transcription" process. In this process, circular plasmid DNA is not purified after enzymatic cleavage; instead, it is directly used for in vitro transcription, resulting in a high-quality mRNA stock solution. This entire process, while ensuring product and process quality, shortens the duration of the existing process.

Process advantages:

  1. Process optimization, reducing IVT steps for simpler operation.
  2. Lower material costs, eliminating the need for post-cleavage purification and quality inspection.
  3. Maintaining mRNA quality and yield.


1. Yields and Integrity:

Using the 1μg of linearized 2K, 4K, and 9K plasmids, the onepot process can generate 150-200μg of mRNA.

Length Yields  Integrity
2K 200 μg 94.00 %
4K 185 μg 92.20 %
9K 150 μg 87.50 %


Integrity testing was performed using capillary electrophoresis (CE) to assess the integrity of fragments with lengths of 2K, 4K, and 9K. The integrity of 2K and 4K fragments was >92%, and the integrity of 9K fragments was >87%.

2. mRNA Capping Efficiency Detection

LC-MS was employed to assess the capping efficiency of the 2K sequence, revealing a capping efficiency of 99.5%.

Top image: HPLC UV chromatogram

Bottom image: Deconvoluted molecular weight spectrum

3. mRNA Polyadenylation Efficiency Detection

LC-MS was used to detect the polyadenylation efficiency of samples, showing results distributed normally.

Top image: Liquid chromatography TIC chromatogram

Bottom image: Deconvoluted molecular weight spectrum

4. mRNA Expression Assay

Transfection of 293T cells with mRNA products synthesized using both the conventional process (Left, linearized plasmids with purification) and the one-pot process (Right, linearized plasmids without purification) showed no difference in fluorescence protein expression after 24 hours of culture.

Ordering Information

Product name SKU Specifications
T7 High Yield RNA Synthesis Kit 10623ES 50/100/500 T
T7 RNA Polymerase GMP-grade (50 U/μL)  10624ES 5000/50000 U
T7 RNA Polymerase GMP-grade(250 U/μL)  10625ES 10/100 KU
10×Transcription Buffer 2 GMP-grade 10670ES 1/10 mL
Pyrophosphatase,Inorganic GMP-grade 0.1 U/μL)  10672ES 10/100/1000 U
Murine RNase inhibitor GMP-grade 10621ES 10/20/100 KU
BspQI GMP-grade  10664ES 500/2500 U
DNase I GMP-grade  10611ES 500/2000/10000 U
mRNA Vaccinia Capping Enzyme GMP-grade  10614ES 2000/10000/100000 U
mRNA Cap 2'-O-Methyltransferase GMP-grade  10612ES 2000/10000/50000 U
10×Capping buffer GMP-grade 10666ES 1/10 mL
S-adenosylmethionine (SAM)(32 mM) 10619ES 0.5/25/500 mL
Pseudouridine-5-triphosphate,trisodium salt solution (100 mM) 10650ES 20 μL/100 μL/1 mL
N1-Me-Pseudo UTP sodium solution(100 mM) 10651ES 20 μL/100 μL/1 mL
ATP Solution(100 mM) 10129ES 1/25/500 mL
CTP Solution(100 mM) 10130ES 1/25/500 mL
UTP Solution(100 mM) 10131ES 1/25/500 mL
GTP Solution(100 mM) 10132ES 1/25/500 mL
NTP Set Solution (ATP, CTP, UTP, GTP, 100 mM each) 10133ES 1 Set (4 vial)
Hieff NGS™ RNA Cleaner 12602ES 1/5/60/450 mL
ATP Tris Solution GMP-grade ( 100 mM) 10652ES 1/5/25/500 ml
CTP Tris Solution GMP-grade ( 100 mM) 10653ES 1/5/25/500 ml
GTP Tris Solution GMP-grade ( 100 mM) 10655ES 1/5/25/500 ml
Pseudo UTP Tris Solution GMP-grade ( 100 mM) 10656ES 1/5/25/500 ml
N1-Me-Pseudo UTP Tris Solution GMP-grade ( 100 mM) 10657ES 1/5/25/500 ml
ARCA (Anti Reverse Cap Analog)  10681ES 1/5/25/500 ml
Double-stranded RNA (dsRNA) ELISA kit  36717ES 48T/96T


Mrna vaccineMrna vaccine enzymes