Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.08.22.671735

Genomic Insights into Wolbachia Strain wCin2USA1 Reveal Promising Cytoplasmic Incompatibility Potential and Next-Generation Dengue Biocontrol

Abstract

Dengue fever poses a growing public health challenge globally, resulting in significant morbidity and economic burden. The use of Wolbachia-mediated biocontrol represents a promising, cost-effective, and environmentally sustainable strategy for managing dengue transmission. However, the susceptibility of existing Wolbachia strains utilized in controlling Aedes aegypti necessitates the investigation of novel strains to enhance dengue control efficacy. This study aimed to identify potential alternative Wolbachia strains for dengue control by comparing the genomes of seven Wolbachia pipientis strains: wMel, wAlbB, wlrr, wHm-C, wAnm, ant7, and a strain isolated from Aedes aegypti. We conducted comprehensive genomic analyses, including phylogenetic assessments, metabolic pathway evaluations, and characterization of the Cytoplasmic Incompatibility Factor (Cif) genes. Our analyses identified the strain wCin2USA1 as a strong candidate for alternative dengue control strategies. This strain demonstrated remarkable genomic similarities to wMel, an already established strain used as biocontrol for Aedes aegypti. Importantly, this strain presented two distinct pairs of Cif genes from different monophyletic types, each homologous to the Cif genes found in wMel and wAlbB. This genetic architecture suggests a high degree of compatibility and demonstrates promising potential for the suppression of Aedes aegypti populations through the induction of Cytoplasmic Incompatibility. The presence of multiple intact prophage regions also suggests greater adaptability compared to established strains. Our findings support the hypothesis that wCin2USA1 could serve as an effective biocontrol agent against dengue transmission. This work provides critical insights into developing innovative Wolbachia-based interventions aimed at mitigating the persistent threat posed by dengue fever. Future research should concentrate on optimizing release methodologies, evaluating ecological impacts, and assessing the strains effectiveness against the dengue virus. Author SummaryDengue fever is one of the fastest-growing mosquito-borne diseases in the world, causing illness and economic challenges in many countries. Current mosquito control methods, such as insecticides, are often costly, less effective over time, and harmful to the environment. An alternative approach uses naturally occurring bacteria called Wolbachia, which live inside insects. When certain Wolbachia strains are introduced into mosquitoes, they can reduce the insects ability to spread viruses like dengue. In our study, we compared the genetic makeup of several Wolbachia strains to identify new candidates that may work better for controlling dengue. We discovered that a strain called wCin2USA1 has strong potential because it shares important features with two strains already used successfully, while also having unique advantages. These include genes that help prevent mosquitoes from reproducing normally when carrying different Wolbachia strains, which can reduce mosquito populations. Our findings suggest that wCin2USA1 could be developed as a new, environmentally friendly tool to help reduce dengue transmission.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Zaeem, I., Jewel, N. A., Rolin, M. H., Karim, D., Akter, A., Mondal, S. I.. 2025-08-26. Genomic Insights into Wolbachia Strain wCin2USA1 Reveal Promising Cytoplasmic Incompatibility Potential and Next-Generation Dengue Biocontrol. https://doi.org/10.1101/2025.08.22.671735

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

PfPHAST: Plasmodium falciparum Public Health Amplicon Sequencing Tool, a Streamlined Panel for Malaria Genomic Surveillance

Genomic tools can support malaria control policy through surveillance of Plasmodium falciparum populations, tracking antimalarial drug resistance, pfhrp2/3 deletions that compromise rapid diagnostic tests, and selection at the circumsporozoite protein (PfCSP) vaccine target, as well as through molecular correction of therapeutic efficacy studies (TES). Multiplex Amplicons for Drug, Diagnostic, Diversity, and Differentiation Haplotypes using Targeted Resequencing (MAD4HatTeR), a comprehensive amplicon sequencing panel covering up to 276 targets, supports these applications but is tailored to research rather than routine programmatic use. We developed P. falciparum Public Health Amplicon Sequencing Tool (PfPHAST), a 56-target derivative of MAD4HatTeR spanning drug resistance loci, pfhrp2/3 deletion, PfCSP genotyping, non-falciparum species identification, and 20 high-heterozygosity microhaplotype loci for TES classification. We compared PfPHAST and MAD4HatTeR using laboratory strain controls, including two-strain dilution series and a five-strain mixture, across parasite densities of 100 to 10,000 parasites/L. At matched per-target depth, PfPHAST achieved a higher quality-control pass rate than MAD4HatTeR (94.4% versus 90.0%) and distributed reads more evenly across targets. The panels showed comparable recall and precision for drug resistance codons and microhaplotypes, reaching near-complete recall above 40% within-sample allele frequency (WSAF) at all densities, with reduced sensitivity for minor alleles below 10% WSAF at low parasite density in both panels. Observed and expected WSAF correlated strongly for both panels, and both resolved a five-strain polyclonal mixture, including a 5% minor strain. By concentrating sequencing capacity on targets of greatest programmatic relevance, PfPHAST offers a scalable, lower-cost alternative to comprehensive research panels without sacrificing performance on shared targets, complementing MAD4HatTeR for routine molecular malaria surveillance.

genomics↗

Structural variation in repeat elements is widespread in normal human tissues and in tumorigenesis

Somatic mosaicism contributes to genomic variation, yet postzygotic structural variants remain under-characterized. We performed long- and short-read WGS from multiple individuals (n=47 normal tissues; n=168 samples) and identified mosaic structural variants in all individuals and germ layers, impacting a median 285.2 kb/genome. Nearly half of breakpoints were independently validated, with tissue distributions reflecting both early and late developmental origins. Most mosaic variants were repeat-mediated and 8.3% overlapped functional elements, an enrichment compared to germline variants. To extend these analyses in samples where long-read sequencing is infeasible, we measured repeat alterations from short-read sequencing, recapitulating mosaic tissue-specific differences. We characterized tumor- and tissue- specific variation in repeats across 15 cancer types and found tumor-related repeat variation to be similar in scale to that of normal mosaic variation. Tracking repeat changes in cell-free DNA provided a noninvasive approach for tumor monitoring. Our analyses revealed widespread repeat-driven structural variation in health and disease.

genomics↗

RNA isoform-resolved multiplexed sequencing with bioorthogonal barcoding

RNA isoform dysregulation drives disease pathogenesis and is the target of FDA-approved splice-switching therapeutics. However, multiplexed sequencing methods discard splice junction information because only 3' termini are barcoded and counted. Here, we repurpose acylation and click chemistries to conjugate bioorthogonal barcodes (bobcodes) directly onto multiple internal positions along cellular RNAs. Bobcoded RNAs from multiple samples are pooled for multiplexed cDNA synthesis, during which reverse transcriptase switches from each RNA template onto its tethered bobcode with greater than 99% accuracy in species mixing experiments. Bobcode attachment intervals set cDNA insert sizes without a library fragmentation step, and priming with poly(dT) or random hexamers selects between 3'-end counting and full-length isoform capture. A bioorthogonal barcode-sequencing (BOB-seq v0.1) drug screen identifies transcriptome-wide on- and off-target RNA splicing effects and outperforms existing multiplexing RNA sequencing methods in workflow simplicity, sample-to-sample variability, and barcoding accuracy. Bobcodes add isoform resolution to scalable multiplexed RNA sequencing.

genomics↗