Across biological scales, gene-regulatory networks employ autorepression (negative feedback) to maintain homeostasis and minimize failure from aberrant expression. Here, we present a proof of concept that disrupting transcriptional negative feedback dysregulates viral gene expression to therapeutically inhibit replication and confers a high evolutionary barrier to resistance. We find that nucleic-acid decoys mimicking cis-regulatory sites act as “feedback disruptors,” break homeostasis, and increase viral transcription factors to cytotoxic levels (termed “open-loop lethality”). Feedback disruptors against herpesviruses reduced viral replication >2-logs without activating innate immunity, showed sub-nM IC₅₀, synergized with standard-of-care antivirals, and inhibited virus replication in mice. In contrast to approved antivirals where resistance rapidly emerged, no feedback-disruptor escape mutants evolved in long-term cultures. For SARS-CoV-2, disruption of a putative feedback circuit also generated open-loop lethality, reducing viral titers by >1-log. These results demonstrate that generating open-loop lethality, via negative-feedback disruption, may yield a class of antimicrobials with a high genetic barrier to resistance.

在生物尺度上，基因调控网络采用自抑制（负反馈）来维持体内平衡并最大限度地减少异常表达引起的失败。在这里，我们提出了一个概念证明，即破坏转录负反馈会失调病毒基因表达，从而在治疗上抑制复制，并赋予抵抗力的高度进化障碍。我们发现模仿顺式调节位点的核酸诱饵充当“反馈干扰器”，打破稳态，并将病毒转录因子增加到细胞毒性水平（称为“开环杀伤力”）。针对疱疹病毒的反馈干扰剂减少了病毒复制 > 2 对数而不激活先天免疫，显示出亚纳米 IC ₅₀，与标准护理抗病毒药物协同作用，并抑制小鼠体内的病毒复制。与迅速出现耐药性的批准抗病毒药物相比，在长期培养中没有进化出反馈干扰逃逸突变体。对于 SARS-CoV-2，假定反馈回路的中断也会产生开环杀伤力，将病毒滴度降低 >1-log。这些结果表明，通过负反馈中断产生开环杀伤力，可能会产生一类具有高抗性遗传屏障的抗菌剂。