Regulated control of virus replication by 4-hydroxytamoxifen-induced splicing

Zhenghao Zhao; Busen Wang; Shipo Wu; Zhe Zhang; Yi Chen; Jinlong Zhang; Yudong Wang; Danni Zhu; Yao Li; Jinghan Xu; Lihua Hou; Wei Chen

doi:10.3389/fmicb.2023.1112580

Regulated control of virus replication by 4-hydroxytamoxifen-induced splicing

Front Microbiol. 2023 Mar 13:14:1112580. doi: 10.3389/fmicb.2023.1112580. eCollection 2023.

Authors

Zhenghao Zhao¹, Busen Wang¹, Shipo Wu¹, Zhe Zhang¹, Yi Chen¹, Jinlong Zhang¹, Yudong Wang¹, Danni Zhu^{1

2}, Yao Li¹, Jinghan Xu¹, Lihua Hou¹, Wei Chen¹

Affiliations

¹ Beijing Institute of Biotechnology, Beijing, China.
² Qingdao Special Servicemen Recuperation Center of PLA Navy, Qingdao, Shandong, China.

Abstract

Designing a modified virus that can be controlled to replicate will facilitate the study of pathogenic mechanisms of virus and virus-host interactions. Here, we report a universal switch element that enables precise control of virus replication after exposure to a small molecule. Inteins mediate a traceless protein splicing-ligation process, and we generate a series of modified vesicular stomatitis virus (VSV) with intein insertion into the nucleocapsid, phosphoprotein, or large RNA-dependent RNA polymerase of VSV. Two recombinant VSV, LC599 and LY1744, were screened for intein insertion in the large RNA-dependent RNA polymerase of VSV, and their replication was regulated in a dose-dependent manner with the small molecule 4-hydroxytamoxifen, which induces intein splicing to restore the VSV replication. Furthermore, in the presence of 4-hydroxytamoxifen, the intein-modified VSV LC599 replicated efficiently in an animal model like a prototype of VSV. Thus, we present a simple and highly adaptable tool for regulating virus replication.

Keywords: intein; post-translation regulation; small molecule switch; vesicular stomatitis virus; virus regulation.