Journal: Nature Communications

Article Title: Small RNA genomics of Aedes aegypti mosquitoes discovers infectious viruses that trigger an RNA interference response

doi: 10.1038/s41467-026-71964-1

Figure Lengend Snippet: a Overview of the Mosquito Small RNA Genomics (MSRG) pipeline applied to a survey of whole mosquitoes from Americas, Asia, Africa and laboratory strains. b Implementation of the VirusDetect program with updated GBVRL and custom databases for comprehensive mosquito virus detection. c Summary tabulation of the samples and RNA libraries analyzed in this study.

Article Snippet: Small RNA libraries were made using NEBNext Small RNA Library Prep Set (NEB #E7330) with up to 5 μg of RNA input.

Techniques: Virus

Journal: Nature Communications

Article Title: Small RNA genomics of Aedes aegypti mosquitoes discovers infectious viruses that trigger an RNA interference response

doi: 10.1038/s41467-026-71964-1

Figure Lengend Snippet: a Bubble plot of vsmRNAs from Africa Ae. aegypti colony strains. Number of reads per million is reflected by bubble diameter, and color represents strand bias of reads, red is plus strand biased, blue is minus strand biased. Dashed pink line boxes mark the PCLV and FORMV noted in panels ( c ) and ( d ), respectively. b Map of Africa locations where the Ae. aegypti colonies or samples originated. Map data ©2025 Google. c Coverage plots of PCLV small RNAs from a selection of African Ae. aegypti showing high M-fragment piRNAs rivaling the S-fragment piRNAs. d The FORMV vsmRNA coverage from African Ae. aegypti colonies from the McBride lab and an independent Kedougou, Senegal sample from Olmo et al. . The black arrow points to male-specific viral piRNA species. e Three examples of FORMV long RNAs sequenced from matched samples in ( d ).

Article Snippet: Small RNA libraries were made using NEBNext Small RNA Library Prep Set (NEB #E7330) with up to 5 μg of RNA input.

Techniques: Selection

Journal: Nature Communications

Article Title: Small RNA genomics of Aedes aegypti mosquitoes discovers infectious viruses that trigger an RNA interference response

doi: 10.1038/s41467-026-71964-1

Figure Lengend Snippet: a Bubble plot of vsmRNAs from lab strains. Reads per million represented by bubble diameter, strand bias represented by color, red is plus strand biased, blue is minus strand biased. See Supplementary Data for sample details. Lab initials: BZL=Benzon Research, MY = M. Younger, DB = D. Brackney, JM = J. Marques, ZT = Z. Tu, GH = G. Hughes, TC = T. Colpitts, BH = B. Hay, GP = G. Pijlman labs. b Coverage plots of long RNAs compared to small RNAs for viruses from the BZL strain of Ae. aegypti females and dormant eggs. c Coverage plots of TMBTLV small RNAs from GP lab strains also infected with Zika virus (ZIKV). d Scatterplot comparing matched small and long RNA libraries from Ae. aegypti . Sequencing RPM are plotted on a logarithmic scale. Sample dots are colored by sex, and clustered samples are in labeled ovals. e Coverage plots of two Florida exhibiting abundant long RNA signal for the Toti-like virus but negligible vsmRNAs in the upper plots that contrast both long and small RNAs against an R1-Ele4 TE. f Coverage plots of long RNAs versus small RNAs for the Formosus virus and R1-Ele4 TE from both males and females of the ENT African colony.

Article Snippet: Small RNA libraries were made using NEBNext Small RNA Library Prep Set (NEB #E7330) with up to 5 μg of RNA input.

Techniques: Infection, Virus, Sequencing, Labeling

Journal: Nature Communications

Article Title: Small RNA genomics of Aedes aegypti mosquitoes discovers infectious viruses that trigger an RNA interference response

doi: 10.1038/s41467-026-71964-1

Figure Lengend Snippet: a Our methodology to molecularly validate the small RNA detection of ISVs are true viruses that can be isolated and verified for triggering the RNAi response in mosquito cells. b RT-PCR detection of TMBTLV RNAs S1 and S2 during multiple rounds of blind passaging, starting with BZL mosquito homogenate as the initial virus infection source placed onto C6/36-NL and Aag2 mosquito cells. The ladder is the 1KbPlus DNA ladder, and uncropped gels are in the source data files. c Virus infection kinetics measured in C6/36-NL and Aag2 cells over 12 days using droplet digital PCR (ddPCR). Flasks with 1 million cells were infected on Day 0 with 20 K viral copies per infection. Virus stocks are from filtered media from subsequent passage from the experiment in (a). Error bars correspond to the 95% confidence interval from Poisson Distribution in the ddPCR analysis algorithm centered around the mean from each reading that contained >15 K droplets replicates. Additional virus infection kinetics measurements are shown in Supplementary Fig. . T-flask illustration from NIAID NIH BioArt Source (bioart.niaid.nih.gov/bioart/303).

Article Snippet: Small RNA libraries were made using NEBNext Small RNA Library Prep Set (NEB #E7330) with up to 5 μg of RNA input.

Techniques: RNA Detection, Isolation, Reverse Transcription Polymerase Chain Reaction, Passaging, Virus, Infection, Digital PCR