mRNA produced by VSW-3 RNAP has high-level translation efficiency with low inflammatory stimulation

Guoquan Wang; Rui Cheng; Qiubing Chen; Yuandong Xu; Bingbing Yu; Bin Zhu; Hao Yin; Heng Xia

doi:10.1016/j.cellin.2022.100056

mRNA produced by VSW-3 RNAP has high-level translation efficiency with low inflammatory stimulation

Cell Insight. 2022 Sep 6;1(5):100056. doi: 10.1016/j.cellin.2022.100056. eCollection 2022 Oct.

Authors

Guoquan Wang¹, Rui Cheng², Qiubing Chen¹, Yuandong Xu³, Bingbing Yu², Bin Zhu², Hao Yin^{1

4

5

6

7

8}, Heng Xia^{1

2

3}

Affiliations

¹ Department of Urology, Frontier Science Centre for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
² Key Laboratory of Molecular Biophysics, The Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
³ Scientific Research Center, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.
⁴ TaiKang Centre for Life and Medical Sciences, TaiKang Medical School, Wuhan University, Wuhan, China.
⁵ Department of Pulmonary and Critical Care Medicine, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
⁶ Department of Pathology, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
⁷ RNA Institute, Wuhan University, Wuhan, China.
⁸ Wuhan Research Centre for Infectious Diseases and Cancer, Chinese Academy of Medical Sciences, Wuhan, China.

Abstract

In vitro preparation of mRNA is a key step for mRNA therapeutics. The widely used T7 RNA polymerase (RNAP) was shown to have many by-products during in vitro transcription (IVT) process, among which double-stranded RNA (dsRNA) is the major by-product to activate the intracellular immune response. Here, we describe the use of a new VSW-3 RNAP that reduced dsRNA production during IVT and the resulting mRNA exhibited low inflammatory stimulation in cells. Compared to T7 RNAP transcripts, these mRNA exhibited superior protein expression levels, with an average of 14-fold increase in Hela cells and 5-fold increase in mice. In addition, we found that VSW-3 RNAP did not require modified nucleotides to improve protein production of IVT products. Our data suggest that VSW-3 RNAP could be a useful tool for mRNA therapeutics.