Imperial College London
Alternate

ProfessorPaiviOjala

Faculty of Medicine, Department of Infectious Disease

Visiting Professor
 
 
 
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Contact

 

+44 (0)20 7594 3971p.ojala Website

 
 
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Location

 

443Medical SchoolSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Elbasani:2020:10.3390/v12090952,
author = {Elbasani, E and Falasco, F and Gramolelli, S and Nurminen, V and Günther, T and Weltner, J and Balboa, D and Grundhoff, A and Otonkoski, T and Ojala, PM},
doi = {10.3390/v12090952},
journal = {Viruses},
title = {Kaposi's Sarcoma-Associated Herpesvirus Reactivation by Targeting of a dCas9-Based Transcription Activator to the ORF50 Promoter},
url = {http://dx.doi.org/10.3390/v12090952},
volume = {12},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - CRISPR activation (CRISPRa) has revealed great potential as a tool to modulate the expression of targeted cellular genes. Here, we successfully applied the CRISPRa system to trigger the Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation in latently infected cells by selectively activating ORF50 gene directly from the virus genome. We found that a nuclease-deficient Cas9 (dCas9) fused to a destabilization domain (DD) and 12 copies of the VP16 activation domain (VP192) triggered a more efficient KSHV lytic cycle and virus production when guided to two different sites on the ORF50 promoter, instead of only a single site. To our surprise, the virus reactivation induced by binding of the stable DD-dCas9-VP192 on the ORF50 promoter was even more efficient than reactivation induced by ectopic expression of ORF50. This suggests that recruitment of additional transcriptional activators to the ORF50 promoter, in addition to ORF50 itself, are needed for the efficient virus production. Further, we show that CRISPRa can be applied to selectively express the early lytic gene, ORF57, without disturbing the viral latency. Therefore, CRISPRa-based systems can be utilized to facilitate virus-host interaction studies by controlling the expression of not only cellular but also of specific KSHV genes.
AU  - Elbasani,E
AU  - Falasco,F
AU  - Gramolelli,S
AU  - Nurminen,V
AU  - Günther,T
AU  - Weltner,J
AU  - Balboa,D
AU  - Grundhoff,A
AU  - Otonkoski,T
AU  - Ojala,PM
DO  - 10.3390/v12090952
PY  - 2020///
SN  - 1999-4915
TI  - Kaposi's Sarcoma-Associated Herpesvirus Reactivation by Targeting of a dCas9-Based Transcription Activator to the ORF50 Promoter
T2  - Viruses
UR  - http://dx.doi.org/10.3390/v12090952
UR  - https://www.ncbi.nlm.nih.gov/pubmed/32867368
UR  - https://www.mdpi.com/1999-4915/12/9/952
UR  - http://hdl.handle.net/10044/1/84664
VL  - 12
ER  -
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