Optimization of In Vitro Transcription for mRNA Production
Millicent Lin, R.K. DeLong, Mathew Brier, Manish Tandon, Li-wen Wang, Yun Chen, Radouane Zouaoui, Aviva Joseph, Eli Kraus and Gregg Nyberg
In vitro transcription (IVT) is a biochemical reaction used for the enzyme-catalyzed synthesis of therapeutic RNA macromolecules, which are generally longer than 200 ribonucleotides and not readily synthesized by automated solid support. Reactions are buffered near neutrality with cations included to stabilize the mRNA and its interaction to enzyme. The T7 RNA polymerase has become the industry standard, and a typical reaction occurs at 37 degrees Celsius for several hours.