Advancing Marburg virus antiviral screening: Optimization of a novel T7 polymerase-independent minigenome system.

Antiviral Research
Bert VanmechelenP. Maes

Abstract

Marburg virus (MARV) is the only known pathogenic filovirus not belonging to the genus Ebolavirus. Minigenomes have proven a useful tool to study MARV, but all existing MARV minigenomes are dependent on the addition of an exogenous T7 RNA polymerase to drive minigenome expression. However, exogenous expression of a T7 polymerase is not always feasible and can act as a confounding factor in compound screening assays. We have developed an alternative minigenome that is controlled by the natively expressed RNA polymerase II. We demonstrate here the characteristics of this new system and its applicability in a wide range of cell types. Our system shows a clear concentration-dependent activity and shows comparable activity to the existing T7 polymerase-based system at higher concentrations, also in difficult-to-transfect cell lines. In addition, we show that our system can be used for compound screening in a 96-well format, thereby providing an attractive alternative to previously developed MARV minigenomes.

References

Nov 11, 1989·Nucleic Acids Research·A LieberM Strauss
Nov 25, 1967·Lancet·C E SmithI Zlotnik
Dec 22, 1967·Deutsche medizinische Wochenschrift·R SiegertG Müller
Mar 12, 1999·The Journal of General Virology·V E VolchkovH Feldmann
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Mar 23, 2002·Cell·Nancy A Woychik, Michael Hampsey
Feb 25, 2005·FEMS Microbiology Letters·Pramila Walpita, Ramon Flick
Dec 28, 2005·Journal of Virology·Sven EnterleinElke Mühlberger
Aug 4, 2009·PLoS Pathogens·Jonathan S TownerPierre E Rollin
Jan 15, 2010·The Journal of General Virology·Jörg WenigenrathStephan Becker
Mar 27, 2015·The Journal of Infectious Diseases·Yoshimi TsudaHideki Ebihara
Mar 7, 2017·Scientific Reports·Michael K LoChristina F Spiropoulou
Jul 1, 2017·Science Translational Medicine·Timothy P SheahanRalph S Baric
May 16, 2019·Archives of Virology·Gaya K AmarasingheJens H Kuhn
Dec 7, 2019·Journal of Autoimmunity·Manuel RojasJuan-Manuel Anaya

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.