Optimized P2A for Reporter Gene Insertion into Nipah Virus Results in Efficient Ribosomal Skipping and Wild-type Lethality

Overview

Journal J Gen Virol

Specialty Microbiology

Date 2016 Jan 20

PMID 26781134

Citations 12

Authors

Arnold Park

Tatyana Yun

Terence E Hill

Tetsuro Ikegami

Terry L Juelich

Jennifer K Smith

Lihong Zhang

Alexander N Freiberg

Benhur Lee

Affiliations

Soon will be listed here.

Abstract

Incorporation of reporter genes within virus genomes is an indispensable tool for interrogation of virus biology and pathogenesis. In previous work, we incorporated a fluorophore into a viral ORF by attaching it to the viral gene via a P2A ribosomal skipping sequence. This recombinant Nipah virus, however, was attenuated in vitro relative to WT virus. In this work, we determined that inefficient ribosomal skipping was a major contributing factor to this attenuation. Inserting a GSG linker before the P2A sequence resulted in essentially complete skipping, significantly improved growth in vitro, and WT lethality in vivo. To the best of our knowledge, this represents the first time a recombinant virus of Mononegavirales with integration of a reporter into a viral ORF has been compared with the WT virus in vivo. Incorporating the GSG linker for improved skipping efficiency whenever functionally important is a critical consideration for recombinant virus design.

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