Flavivirus RNA synthesis in vitro

Radhakrishnan Padmanabhan; Ratree Takhampunya; Tadahisa Teramoto; Kyung H. Choi

doi:10.1016/j.ymeth.2015.08.002

Flavivirus RNA synthesis in vitro

Radhakrishnan Padmanabhan, Ratree Takhampunya, Tadahisa Teramoto, Kyung H. Choi

Research output: Contribution to journal › Review article › peer-review

4 Scopus citations

Abstract

Establishment of in vitro systems to study mechanisms of RNA synthesis for positive strand RNA viruses have been very useful in the past and have shed light on the composition of protein and RNA components, optimum conditions, the nature of the products formed, cis-acting RNA elements and trans-acting protein factors required for efficient synthesis. In this review, we summarize our current understanding regarding the requirements for flavivirus RNA synthesis in vitro. We describe details of reaction conditions, the specificity of template used by either the multi-component membrane-bound viral replicase complex or by purified, recombinant RNA-dependent RNA polymerase. We also discuss future perspectives to extend the boundaries of our knowledge.

Original language	English (US)
Pages (from-to)	20-34
Number of pages	15
Journal	Methods
Volume	91
DOIs	https://doi.org/10.1016/j.ymeth.2015.08.002
State	Published - 2015

Keywords

Cytoplasmic membrane-bound viral replication complex
Endogenous and exogenous viral RNA templates
Heparin trap
Polarity of RNA product by RNase H mapping
RNA-dependent RNA polymerase from positive strand mosquito-borne Flaviviridae
Temperature-dependent de novo initiation and elongation

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology

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10.1016/j.ymeth.2015.08.002

Cite this

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title = "Flavivirus RNA synthesis in vitro",

abstract = "Establishment of in vitro systems to study mechanisms of RNA synthesis for positive strand RNA viruses have been very useful in the past and have shed light on the composition of protein and RNA components, optimum conditions, the nature of the products formed, cis-acting RNA elements and trans-acting protein factors required for efficient synthesis. In this review, we summarize our current understanding regarding the requirements for flavivirus RNA synthesis in vitro. We describe details of reaction conditions, the specificity of template used by either the multi-component membrane-bound viral replicase complex or by purified, recombinant RNA-dependent RNA polymerase. We also discuss future perspectives to extend the boundaries of our knowledge.",

keywords = "Cytoplasmic membrane-bound viral replication complex, Endogenous and exogenous viral RNA templates, Heparin trap, Polarity of RNA product by RNase H mapping, RNA-dependent RNA polymerase from positive strand mosquito-borne Flaviviridae, Temperature-dependent de novo initiation and elongation",

author = "Radhakrishnan Padmanabhan and Ratree Takhampunya and Tadahisa Teramoto and Choi, {Kyung H.}",

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year = "2015",

doi = "10.1016/j.ymeth.2015.08.002",

language = "English (US)",

volume = "91",

pages = "20--34",

journal = "Methods",

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T1 - Flavivirus RNA synthesis in vitro

AU - Padmanabhan, Radhakrishnan

AU - Takhampunya, Ratree

AU - Teramoto, Tadahisa

AU - Choi, Kyung H.

PY - 2015

Y1 - 2015

N2 - Establishment of in vitro systems to study mechanisms of RNA synthesis for positive strand RNA viruses have been very useful in the past and have shed light on the composition of protein and RNA components, optimum conditions, the nature of the products formed, cis-acting RNA elements and trans-acting protein factors required for efficient synthesis. In this review, we summarize our current understanding regarding the requirements for flavivirus RNA synthesis in vitro. We describe details of reaction conditions, the specificity of template used by either the multi-component membrane-bound viral replicase complex or by purified, recombinant RNA-dependent RNA polymerase. We also discuss future perspectives to extend the boundaries of our knowledge.

AB - Establishment of in vitro systems to study mechanisms of RNA synthesis for positive strand RNA viruses have been very useful in the past and have shed light on the composition of protein and RNA components, optimum conditions, the nature of the products formed, cis-acting RNA elements and trans-acting protein factors required for efficient synthesis. In this review, we summarize our current understanding regarding the requirements for flavivirus RNA synthesis in vitro. We describe details of reaction conditions, the specificity of template used by either the multi-component membrane-bound viral replicase complex or by purified, recombinant RNA-dependent RNA polymerase. We also discuss future perspectives to extend the boundaries of our knowledge.

KW - Cytoplasmic membrane-bound viral replication complex

KW - Endogenous and exogenous viral RNA templates

KW - Heparin trap

KW - Polarity of RNA product by RNase H mapping

KW - RNA-dependent RNA polymerase from positive strand mosquito-borne Flaviviridae

KW - Temperature-dependent de novo initiation and elongation

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DO - 10.1016/j.ymeth.2015.08.002

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SN - 1046-2023

VL - 91

SP - 20

EP - 34

JO - Methods

JF - Methods

ER -

Flavivirus RNA synthesis in vitro

Abstract

Keywords

ASJC Scopus subject areas

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