人口规模的数据库已扩展到数百万个蛋白质编码变体，但对其机制后果的深入了解却滞后。在这里，我们介绍了 PROD-ATAC，这是一种用于发现蛋白质编码变体对染色质调控影响的高通量方法。汇集的变体文库在与疾病无关的细胞系中表达，并且对转座酶可及的染色质进行单细胞测定可以解决每个变体对染色质景观的影响。使用 PROD-ATAC，我们表征了 100 多种肿瘤融合（致癌嵌合蛋白）和对照的影响，并揭示了染色质重塑对于跨越巨大融合频率范围的融合来说是常见的。此外，融合引起的失调可能与背景无关，因为观察到的机制通常与癌症和细胞类型特异性的先验知识重叠。我们还表明，功能获得活动在肿瘤融合中很常见。这项工作开始勾画出融合引起的染色质改变的全局图。我们认为，不同的肿瘤融合之间可能存在趋同机制，并且肿瘤中不同频率的融合之间可能存在共同的失调模式。 PROD-ATAC 可推广到任何一组蛋白质编码变体。© 2024。作者获得 Springer Nature America, Inc. 的独家许可。

Population-scale databases have expanded to millions of protein-coding variants, yet insight into their mechanistic consequences has lagged. Here we present PROD-ATAC, a high-throughput method for discovering the effects of protein-coding variants on chromatin regulation. A pooled variant library is expressed in a disease-agnostic cell line, and single-cell assay for transposase-accessible chromatin resolves each variant's effect on the chromatin landscape. Using PROD-ATAC, we characterized the effects of more than 100 oncofusions (cancer-causing chimeric proteins) and controls and revealed that chromatin remodeling is common to fusions spanning an enormous range of fusion frequencies. Furthermore, fusion-induced dysregulation can be context agnostic, as observed mechanisms often overlapped with cancer and cell-type-specific prior knowledge. We also showed that gain-of-function activity is common among oncofusions. This work begins to outline a global map of fusion-induced chromatin alterations. We suggest that there might be convergent mechanisms among disparate oncofusions and shared modes of dysregulation among fusions present in tumors at different frequencies. PROD-ATAC is generalizable to any set of protein-coding variants.© 2024. The Author(s), under exclusive licence to Springer Nature America, Inc.