A Reverse Genetics System for Zika Virus Based on a Simple Molecular Cloning Strategy

Viruses
Maximilian MünsterRalf Bartenschlager

Abstract

The Zika virus (ZIKV) has recently attracted major research interest as infection was unexpectedly associated with neurological manifestations in developing foetuses and with Guillain-Barré syndrome in infected adults. Understanding the underlying molecular mechanisms requires reverse genetic systems, which allow manipulation of infectious cDNA clones at will. In the case of flaviviruses, to which ZIKV belongs, several reports have indicated that the construction of full-length cDNA clones is difficult due to toxicity during plasmid amplification in Escherichia coli. Toxicity of flaviviral cDNAs has been linked to the activity of cryptic prokaryotic promoters within the region encoding the structural proteins leading to spurious transcription and expression of toxic viral proteins. Here, we employ an approach based on in silico prediction and mutational silencing of putative promoters to generate full-length cDNA clones of the historical MR766 strain and the contemporary French Polynesian strain H/PF/2013 of ZIKV. While for both strains construction of full-length cDNA clones has failed in the past, we show that our approach generates cDNA clones that are stable on single bacterial plasmids and give rise to infectious viruses w...Continue Reading

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Citations

Oct 23, 2019·Nature Microbiology·Christopher J NeufeldtRalf Bartenschlager
Jun 3, 2020·Antimicrobial Agents and Chemotherapy·Nicholas S EyreMichael R Beard
Nov 6, 2018·Viruses·Ginés Ávila-PérezLuis Martínez-Sobrido
Mar 17, 2019·Viruses·Luis Martinez-Sobrido, Fernando Almazán
Jul 20, 2020·Current Opinion in Virology·Carlos Sandoval-Jaime
Jun 22, 2020·Current Opinion in Virology·Camila R Fontes-GarfiasPei-Yong Shi
Mar 16, 2021·Frontiers in Microbiology·Iasmim Silva de MelloMyrna Cristina Bonaldo
Oct 8, 2021·Nature·Suzanne J F KapteinJohan Neyts
Dec 16, 2021·Nature Communications·Keisuke TabataRalf Bartenschlager
Dec 18, 2018··Clément MazeaudWesley Freppel

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Datasets Mentioned

BETA
KJ776791
DQ859059
KX197192

Methods Mentioned

BETA
in vitro transcription
PCR
electrophoresis
nucleotide exchange
restriction digest
transfection

Software Mentioned

Neural Network

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