【Scientific Research News】Pure mRNA from Shangri-La——Zhu Bin's research group of Huake University discovered a new RNA synthesis tool enzyme

On October 26, 2022, Professor Zhu Bin's team from the School of Life Sciences, Huazhong University of Science and Technology, published a report entitled "Psychrophilic phage VSW-3 RNA polymerase reduces both terminal and full-length dsRNA byproducts in in vitro", a well-known international journal in the field of RNA transcription"
.

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mRNA drugs and vaccines are currently the hot direction of biomedicine, and the only way for mRNA synthesis in vitro is in vitro transcription (IVT)
based on single-subunit RNA polymerase.
In view of the scarcity and research difficulty of single subunit RNA polymerase in nature, in vitro transcription technology tool enzymes are monopolized by bacteriophage T7 RNA polymerase discovered more than 40 years ago, and the performance defects of T7 RNA polymerase itself have become a bottleneck
restricting the development of RNA field.
The most serious problem for mRNA drugs is the double-stranded RNA (dsRNA) byproduct produced by T7 RNA polymerase in vitro transcription, because double-stranded RNA mimics the viral genome to activate cellular immune responses that can trigger strong cytotoxicity
even in trace amounts.
Compared with the many DNA polymerase tools with different performance in in vitro DNA synthesis technology, RNA in vitro synthesis urgently requires the discovery and development
of more new tool enzymes.

Zhu Bin's research group used the explosive growth of environmental microbial genome resources in recent years to report the discovery and development of all new bacteriophage monosubunit RNA polymerases since the new century, including Syn5 RNA polymerase, KP34 RNA polymerase and VSW-3 RNA polymerase, which have formed an international characteristic research direction
.
In this study, VSW-3 RNA polymerase
was reported from the Napa Sea, a glacial lake in the seasonal wetlands of the Shangri-La Plateau in Yunnan.
Due to its characteristics of binding ability to RNA and DNA ends, VSW-3 RNA polymerase effectively reduces the end extension of T7 RNA polymerase with RNA as a template and the transcription initiation of DNA terminals in vitro transcription, so the terminal double-stranded RNA and full-length double-stranded RNA significantly present in T7 product are almost undetectable in VSW-3 product, which greatly improves the purity of in vitro synthetic RNA, especially suitable for RNA applications such as mRNA drugs and vaccines
in vivo 。 At the same time, the low temperature synthesis of VSW-3 RNA polymerase and the insensitivity to the second type of T7 transcription terminator further improved the purity
of RNA synthesis.

Xia Heng, a postdoctoral fellow at the School of Life Sciences, Huazhong University of Science and Technology, and Yu Bingbing, a doctoral student, are the first authors of the paper, Huazhong University of Science and Technology is the first paper, and Zhu Bin, professor of the School of Life Sciences, Huazhong University of Science and Technology, is the author
of the communication.
This research was supported
by the National Natural Science Foundation of China Original Exploration Project and Shenzhen Basic Research Key Project.