Genome editing in archaeal viruses and endogenous viral protein purification

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Archaea-infecting viruses are morphologically and genomically among the most diverse entities. Unfortunately, they are also fairly understudied due to a lack of efficient genetic tools. Here, we present a detailed protocol for the CRISPR/Cas-based genome editing of the virus SIRV2 infecting the genus Sulfolobus, which could easily be adapted to other archaeal viruses. This protocol also includes the procedure for endogenous viral protein purification and identification, allowing for assessing the molecular mechanisms behind virus life cycle and virus-host interactions. For complete details on the use and execution of this protocol, please refer to Mayo-Muñoz et al. (2018) and Bhoobalan-Chitty et al. (2019).

OriginalsprogEngelsk
Artikelnummer100791
TidsskriftSTAR Protocols
Vol/bind2
Udgave nummer4
Antal sider27
ISSN2666-1667
DOI
StatusUdgivet - 2021

Bibliografisk note

Funding Information:
This work was supported by the Danish Council for Independent Research/Technology and Production DFF-7017-00060 (to X.P.), the Novo Nordisk Fonden /Hallas Møller Ascending Investigator Grant NNF17OC0031154 (to X.P.), and the Novo Nordisk Fonden Postdoctoral Fellowship in Bioscience and Basic Biomedicine Grant NNF21OC0067491 (to Y.B.-C.). We thank all the past and present members of the Danish Archaea Centre for their input and contributions and especially Safir Hien Phan for her guidance with preparation of protocols relating to the Sulfolobus medium.

Funding Information:
This work was supported by the Danish Council for Independent Research/Technology and Production DFF-7017-00060 (to X.P.), the Novo Nordisk Fonden/Hallas M?ller Ascending Investigator Grant NNF17OC0031154 (to X.P.), and the Novo Nordisk Fonden Postdoctoral Fellowship in Bioscience and Basic Biomedicine Grant NNF21OC0067491 (to Y.B.-C.). We thank all the past and present members of the Danish Archaea Centre for their input and contributions and especially Safir Hien Phan for her guidance with preparation of protocols relating to the Sulfolobus medium. Y.B.-C. and X.P. conceived the project and designed the experiments, Y.B.-C. and L.A. performed the experiments; L.A. and Y.B.-C. wrote the manuscript together with X.P. All authors read and approved the final manuscript. The authors declare no competing interests.

Publisher Copyright:
© 2021 The Authors

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