anti-CHIKV/DENV Dual Targeting Introns in a Bicistronic Configuration.

Each of the trans-splicing dual targeting introns was tagged downstream of the ΔN Bax 3’ exon (active, further truncated version of tBax) with the mCherry fluorescent marker gene expressed from an IRES sequence of the Drosophila C Virus (DCV). Expression of these constructs was driven by the Drosophila melanogaster Actin 5c promoter. Each of these Dicistrovirus IRES sequences was previously determined to yield the highest levels of expression in Ae. aegypti mosquito cells [48]. The bicistronic configuration allowed monitoring for the presence and expression of these constructs within cell cultures through dual expression of antiviral intron-ΔN Bax and mCherry cistronic portions of the plasmid. Version 1 (v1) and Version 2 (v2) refer to the order of the CHIKV and DENV specific P10 helix sequences. CHIKV = Chikungunya virus targeting sequences; DENV = Dengue virus targeting sequences; IRES = internal ribosome entry site; EGS = external guide sequence; IGS = internal guide sequence; BL = bulge loop; TSD = trans splicing domain; P10 = P10 helix.