Suppression of the arboviruses dengue and chikungunya using a dual-acting group-I intron coupled with conditional expression of the bax C-terminal domain

Autor(es): Carter James R, Taylor Samantha, Fraser Tresa S, Kucharski Cheryl A, Dawson James L, Fraser Malcolm J


Resumo: In portions of South Asia, vectors - patients co-infected with dengue (DENV) - chikungunya (CHIKV) are on the rise, with the potential for this occurrence in other regions of the world, for example the United States. Therefore, we engineered an antiviral approach that suppresses the replication of both arboviruses in mosquito cells using a single antiviral group I intron. We devised unique configurations of internal, external, - guide sequences that permit homologous recognition - splicing with conserved target sequences in the genomes of both viruses using a single trans-splicing Group I intron, - examined their effectiveness to suppress infections of DENV - CHIKV in mosquito cells when coupled with a proapoptotic 3' exon, ?N Bax. RT-PCR demonstrated the utility of these introns in trans-splicing the ?N Bax sequence downstream of either the DENV or CHIKV target site in transformed Aedes albopictus C6/36 cells, independent of the order in which the virus specific targeting sequences were inserted into the construct. This trans-splicing reaction forms DENV or CHIKV ?N Bax RNA fusions that led to apoptotic cell death as evidenced by annexin V staining, caspase, - DNA fragmentation assays. TCID50-IFA analyses demonstrate effective suppression of DENV - CHIKV infections by our anti-arbovirus group I intron approach. This represents the first report of a dual-acting Group I intron, - demonstrates that we can target DENV - CHIKV RNAs in a sequence specific manner with a single, uniquely configured CHIKV/DENV dual targeting group I intron, leading to replication suppression of both arboviruses, - thus providing a promising single antiviral for the transgenic suppression of multiple arboviruses.


Imprenta: PloS One, v. 10, n. 11, p. e0139899, 2015


Identificador do objeto digital: 10.1371/journal.pone.0139899


Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - Cell ; Chikungunya virus - DNA ; Chikungunya virus - RNA ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Dengue


Data de publicação: 2015