Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.omtn.2017.03.004
DC FieldValue
dc.titleExpression of Herpes Simplex Virus Thymidine Kinase/Ganciclovir by RNA Trans-Splicing Induces Selective Killing of HIV-Producing Cells
dc.contributor.authorIngemarsdotter, Carin K
dc.contributor.authorSushmita Poddar
dc.contributor.authorMercier, Sarah
dc.contributor.authorVolker Patzel
dc.contributor.authorLever, Andrew M L
dc.date.accessioned2018-01-24T03:53:40Z
dc.date.available2018-01-24T03:53:40Z
dc.date.issued2017-06-01
dc.identifier.citationIngemarsdotter, Carin K, Sushmita Poddar, Mercier, Sarah, Volker Patzel, Lever, Andrew M L (2017-06-01). Expression of Herpes Simplex Virus Thymidine Kinase/Ganciclovir by RNA Trans-Splicing Induces Selective Killing of HIV-Producing Cells. Molecular Therapy - Nucleic Acids 7 : 140-154. ScholarBank@NUS Repository. https://doi.org/10.1016/j.omtn.2017.03.004
dc.identifier.issn2162-2531
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/138524
dc.description.abstractAntiviral strategies targeting hijacked cellular processes are less easily evaded by the virus than viral targets. If selective for viral functions, they can have a high therapeutic index. We used RNA trans-splicing to deliver the herpes simplex virus thymidine kinase-ganciclovir (HSV-tk/GCV) cell suicide system into HIV-producing cells. Using an extensive in silico bioinformatics and RNA structural analysis approach, ten HIV RNA trans-splicing constructs were designed targeting eight different HIV splice donor or acceptor sites and were tested in cells expressing HIV. Trans-spliced mRNAs were identified in HIV-expressing cells using qRT-PCR with successful detection of fusion RNA transcripts between HIV RNA and the HSV-tk RNA transcripts from six of ten candidate RNA trans-splicing constructs. Conventional PCR and Sanger sequencing confirmed RNA trans-splicing junctions. Measuring cell viability in the presence or absence of GCV expression of HSV-tk by RNA trans-splicing led to selective killing of HIV-producing cells using either 3' exon replacement or 5' exon replacement in the presence of GCV. Five constructs targeting four HIV splice donor and acceptor sites, D4, A5, A7, and A8, involved in regulating the generation of multiple HIV RNA transcripts proved to be effective for trans-splicing mediated selective killing of HIV-infected cells, within which individual constructs targeting D4 and A8 were the most efficient.
dc.language.isoen
dc.publisherElsevier
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectHIV
dc.subjectHSV-tk/GCV
dc.subjectRNA splicing
dc.subjectRNA trans-splicing
dc.subjectexon replacement
dc.subjectgene therapy
dc.subjectherpes simplex virus thymidine kinase-ganciclovir
dc.typeArticle
dc.contributor.departmentMICROBIOLOGY & IMMUNOLOGY
dc.description.doi10.1016/j.omtn.2017.03.004
dc.description.sourcetitleMolecular Therapy - Nucleic Acids
dc.description.volume7
dc.description.page140-154
dc.identifier.isiut000401233700014
dc.published.statePublished
dc.grant.idR-182-000-163-646
dc.grant.fundingagencyNational University of Singapore
Appears in Collections:Staff Publications
Elements

Show simple item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
2017-expression_herpes_simplex_virus_thymidine-pub.pdf2.74 MBAdobe PDF

OPEN

PublishedView/Download

Google ScholarTM

Check

Altmetric


This item is licensed under a Creative Commons License Creative Commons