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Here we show that in over 50 participants, plasma viremia is markedly driven by highly clonal HIV-1 RNA populations carrying defects in the 5\u2032L. Across individuals, dominant clones with 5\u2032L defects clustered around the major splice donor (MSD) accounted for the vast majority of circulating HIV-1 RNA. To enable rapid, scalable profiling, we developed CLAWS (Capturing 5\u2032 Leader Anomalies Without Sequencing), a digital PCR assay that distinguishes intact from defective 5\u2032L RNA. CLAWS recapitulated sequencing-based estimates and detected low-abundance defective RNA early after ART initiation, revealing that defective genomes emerge early and become predominant during long-term therapy. These findings identify 5\u2032L-defective genomes as the predominant driver of NSV and establish CLAWS as a practical tool for monitoring viremia in clinical and cure-related settings.<\/jats:p>","DOI":"10.1038\/s41467-026-73475-5","type":"journal-article","created":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T09:04:21Z","timestamp":1780909461000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["5\u2032 leader defects drive persistent HIV-1 viremia on long-term ART"],"prefix":"10.1038","volume":"17","author":[{"given":"Julia 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