Broad-Spectrum Antiviral Activity of Small Interfering RNA Targeting the Conserved RNA Termini of Lassa Virus

Abstract

Small interfering RNAs targeting the conserved RNA termini upstream of NP and L gene were found to reduce reporter gene expression from Lassa virus replicon and Lassa virus mRNA expression construct and to inhibit replication of different Lassa virus strains, lymphocytic choriomeningitis virus, and Mopeia virus in cell culture.

Reverse Genetics Recovery of Lujo Virus and Role of Virus RNA Secondary Structures in Efficient Virus Growth

“…To obtain a recombinant Lujo virus, we focused our initial efforts on sequencing the genome termini, as these are essential cis elements of viral promoters (6,15,17,30). The sequences of the genomic RNA termini of Pichínde virus (33), Tacaríbe virus (34), and LCMV (14) implied the presence of a nontemplated pppG originating from internal priming by a 5= dinucleotide triphosphate (5= pppGpC) and realigning the primer with the terminal 3= G. In contrast to other arenavirus results, we detected not only a possible nontemplated 5= pppG in Lujo virus but also an additional 3= C at the termini of both segments, albeit a significant proportion of S termini end with 3= G, as in other arenaviruses (1,2,5,14,27,34). Our data suggest that Lujo virus genome termini differ significantly from those of other arenaviruses.…”

“…1A). Interestingly, the Lujo virus L segment 3= end terminated with an extra C following the arenavirus consensus sequence (1,27). Similarly, the terminal 3= nucleotide on the S segment was a mix of a C and A (Fig.…”

“…The circular nature of the viral genome is likely maintained by complementary inverted sequences at the 5= and 3= ends of the viral RNA segments, which form a panhandle-like RNA structure (4,5,11). The 19 terminal nucleotides (nt), which may contribute to formation of the panhandle structure on both segments, are strictly conserved (27) and critical for replication (17,30).…”

Efficient Reverse Genetics Generation of Infectious Junin Viruses Differing in Glycoprotein Processing

“…With regard to the JUNV RNA terminal sequences derived here, it is now clear that all the known arenavirus RNA termini share the feature whereby the termini of the L RNA segment have the potential to form a perfect panhandle structure, but the termini of the S RNA segment contain mismatches that would result in an imperfect structure (30). The strict conservation of this feature suggests it has a likely role in balancing the relative transcription, replication, and packaging efficiency of the S and L RNA segments.…”

Rescue of a Recombinant Machupo Virus from Cloned cDNAs and In Vivo Characterization in Interferon (αβ/γ) Receptor Double Knockout Mice

“…MG systems of other arenaviruses have been proven to be useful tools in dissecting the role of viral proteins in arenavirus RNA replication and transcription, identifying the function of the IGR as the transcrip- tion terminating signal, studying the role of Z as a matrix protein, and mapping the cap snatching domain of L protein (47)(48)(49)(50)(51)(52). The development of an MG for MACV may assist in studying the replication of this virus in the BSL-2 environment, which may also be utilized as a tool for testing antivirals as it has been shown for the Lassa virus MG system (53). Our studies also provided preliminary evidence for the compatibility of Candid #1 L protein and NP for supporting MACV RNA template replication.…”