m6A at the HIV-1 5′ Leader: A Cross-Study Evidence Audit of Genome Packaging, Viral-RNA Dose, and Innate Sensing

This document is an evidence audit, not a report of new experimental data. It compares measurements made in separate published studies and states precisely what can and cannot be inferred from the combined record. Chen et al. (PLoS Pathogens, 2021) reported that N6-methyladenosine (m6A) on HIV-1 RNA suppresses RIG-I-dependent type-I interferon (IFN-I) induction. Pereira-Montecinos et al. (Nucleic Acids Research, 2022) subsequently showed that FTO-mediated demethylation increases incorporation of full-length RNA into viral particles by approximately five-fold per CAp24 equivalent. Because the Chen infection experiments normalised viral inocula by p24, equal-p24 inocula need not contain equal numbers of HIV-1 genomes. The concern is not new: a reviewer of the Chen study explicitly identified pre-challenge viral-RNA quantification as a critical missing control. This audit connects that documented concern to the later independent packaging measurement, distinguishes the experimental arms that are exposed to the problem from those that are not, and explains why the packaging fold-change cannot be converted into a predicted IFN-I effect size in the absence of a measured genome-dose