Effective suppression of Dengue virus using a novel group-I intron that induces apoptotic cell death upon infection through conditional expression of the Bax C-terminal domain

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dc.contributor.author Carter, James R.
dc.contributor.author Keith, James H.
dc.contributor.author Fraser, Tresa S.
dc.contributor.author Dawson, James L.
dc.contributor.author Kucharski, Cheryl A.
dc.contributor.author Horne, Kate M.
dc.contributor.author Higgs, Stephen
dc.contributor.author Fraser, Malcolm J., Jr.
dc.date.accessioned 2014-11-06T19:14:12Z
dc.date.available 2014-11-06T19:14:12Z
dc.date.issued 2014-11-06
dc.identifier.uri http://hdl.handle.net/2097/18649
dc.description.abstract Introduction: Approximately 100 million confirmed infections and 20,000 deaths are caused by Dengue virus (DENV) outbreaks annually. Global warming and rapid dispersal have resulted in DENV epidemics in formally non-endemic regions. Currently no consistently effective preventive measures for DENV exist, prompting development of transgenic and paratransgenic vector control approaches. Production of transgenic mosquitoes refractory for virus infection and/or transmission is contingent upon defining antiviral genes that have low probability for allowing escape mutations, and are equally effective against multiple serotypes. Previously we demonstrated the effectiveness of an anti-viral group I intron targeting U143 of the DENV genome in mediating trans-splicing and expression of a marker gene with the capsid coding domain. In this report we examine the effectiveness of coupling expression of ΔN Bax to trans-splicing U143 intron activity as a means of suppressing DENV infection of mosquito cells. Results: Targeting the conserved DENV circularization sequence (CS) by U143 intron trans-splicing activity appends a 3’ exon RNA encoding ΔN Bax to the capsid coding region of the genomic RNA, resulting in a chimeric protein that induces premature cell death upon infection. TCID50-IFA analyses demonstrate an enhancement of DENV suppression for all DENV serotypes tested over the identical group I intron coupled with the non-apoptotic inducing firefly luciferase as the 3’ exon. These cumulative results confirm the increased effectiveness of this αDENV-U143-ΔN Bax group I intron as a sequence specific antiviral that should be useful for suppression of DENV in transgenic mosquitoes. Annexin V staining, caspase 3 assays, and DNA ladder observations confirm DCA-ΔN Bax fusion protein expression induces apoptotic cell death. Conclusion: This report confirms the relative effectiveness of an anti-DENV group I intron coupled to an apoptosis-inducing ΔN Bax 3’ exon that trans-splices conserved sequences of the 5’ CS region of all DENV serotypes and induces apoptotic cell death upon infection. Our results confirm coupling the targeted ribozyme capabilities of the group I intron with the generation of an apoptosis-inducing transcript increases the effectiveness of infection suppression, improving the prospects of this unique approach as a means of inducing transgenic refractoriness in mosquitoes for all serotypes of this important disease. en_US
dc.language.iso en_US en_US
dc.relation.uri http://www.virologyj.com/content/11/1/111 en_US
dc.subject Dengue en_US
dc.subject Trans-splicing en_US
dc.subject Group I intron en_US
dc.subject Ribozyme en_US
dc.subject Mosquito en_US
dc.subject Antiviral en_US
dc.subject Suppression en_US
dc.title Effective suppression of Dengue virus using a novel group-I intron that induces apoptotic cell death upon infection through conditional expression of the Bax C-terminal domain en_US
dc.type Article (publisher version) en_US
dc.date.published 2014 en_US
dc.citation.doi doi:10.1186/1743-422X-11-111 en_US
dc.citation.jtitle Virology Journal en_US
dc.citation.spage 111 en_US
dc.citation.volume 11 en_US
dc.contributor.authoreid kmhorne en_US
dc.contributor.authoreid shiggs en_US

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