From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging.
Identifieur interne : 000115 ( France/Analysis ); précédent : 000114; suivant : 000116From Cells to Virus Particles: Quantitative Methods to Monitor RNA Packaging.
Auteurs : Mireia Ferrer [France] ; Simon Henriet [Norvège] ; Célia Chamontin [France] ; Sébastien Lainé [France] ; Marylène Mougel [France]Source :
- Viruses [ 1999-4915 ] ; 2016.
Descripteurs français
- KwdFr :
- MESH :
- métabolisme : ARN viral, Virion.
- physiologie : Retroviridae.
- Animaux, Assemblage viral, Biologie moléculaire, Humains, Virologie.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : RNA, Viral.
- metabolism : Virion.
- methods : Molecular Biology, Virology.
- physiology : Retroviridae.
- Animals, Humans, Virus Assembly.
Abstract
In cells, positive strand RNA viruses, such as Retroviridae, must selectively recognize their full-length RNA genome among abundant cellular RNAs to assemble and release particles. How viruses coordinate the intracellular trafficking of both RNA and protein components to the assembly sites of infectious particles at the cell surface remains a long-standing question. The mechanisms ensuring packaging of genomic RNA are essential for viral infectivity. Since RNA packaging impacts on several essential functions of retroviral replication such as RNA dimerization, translation and recombination events, there are many studies that require the determination of RNA packaging efficiency and/or RNA packaging ability. Studies of RNA encapsidation rely upon techniques for the identification and quantification of RNA species packaged by the virus. This review focuses on the different approaches available to monitor RNA packaging: Northern blot analysis, ribonuclease protection assay and quantitative reverse transcriptase-coupled polymerase chain reaction as well as the most recent RNA imaging and sequencing technologies. Advantages, disadvantages and limitations of these approaches will be discussed in order to help the investigator to choose the most appropriate technique. Although the review was written with the prototypic simple murine leukemia virus (MLV) and complex human immunodeficiency virus type 1 (HIV-1) in mind, the techniques were described in order to benefit to a larger community.
DOI: 10.3390/v8080239
PubMed: 27556480
Affiliations:
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pubmed:27556480Le document en format XML
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How viruses coordinate the intracellular trafficking of both RNA and protein components to the assembly sites of infectious particles at the cell surface remains a long-standing question. The mechanisms ensuring packaging of genomic RNA are essential for viral infectivity. Since RNA packaging impacts on several essential functions of retroviral replication such as RNA dimerization, translation and recombination events, there are many studies that require the determination of RNA packaging efficiency and/or RNA packaging ability. Studies of RNA encapsidation rely upon techniques for the identification and quantification of RNA species packaged by the virus. This review focuses on the different approaches available to monitor RNA packaging: Northern blot analysis, ribonuclease protection assay and quantitative reverse transcriptase-coupled polymerase chain reaction as well as the most recent RNA imaging and sequencing technologies. Advantages, disadvantages and limitations of these approaches will be discussed in order to help the investigator to choose the most appropriate technique. Although the review was written with the prototypic simple murine leukemia virus (MLV) and complex human immunodeficiency virus type 1 (HIV-1) in mind, the techniques were described in order to benefit to a larger community.</div></front></TEI><affiliations><list><country><li>France</li><li>Norvège</li></country><region><li>Languedoc-Roussillon</li><li>Occitanie (région administrative)</li></region></list><tree><country name="France"><region name="Occitanie (région administrative)"><name sortKey="Ferrer, Mireia" sort="Ferrer, Mireia" uniqKey="Ferrer M" first="Mireia" last="Ferrer">Mireia Ferrer</name></region><name sortKey="Chamontin, Celia" sort="Chamontin, Celia" uniqKey="Chamontin C" first="Célia" last="Chamontin">Célia Chamontin</name><name sortKey="Laine, Sebastien" sort="Laine, Sebastien" uniqKey="Laine S" first="Sébastien" last="Lainé">Sébastien Lainé</name><name sortKey="Mougel, Marylene" sort="Mougel, Marylene" uniqKey="Mougel M" first="Marylène" last="Mougel">Marylène Mougel</name></country><country name="Norvège"><noRegion><name sortKey="Henriet, Simon" sort="Henriet, Simon" uniqKey="Henriet S" first="Simon" last="Henriet">Simon Henriet</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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