联合生物能源研究所Patrick M. Shih研究小组近日取得一项新成果。经过不懈努力，他们提出了植物细胞中转录调控因子的多重分析。相关论文发表在2025年11月25日出版的《自然—生物技术》杂志上。

在这里，该课题组介绍了利用测序富集植物核反式元件报告基因的方法（ENTRAP-seq），这是一种高通量的方法，将蛋白质编码文库引入植物细胞中，以驱动基于核磁分选的报告基因，从而能够多重测量蛋白质变体的调节活性。使用ENTRAP-seq和机器学习，该研究团队筛选了1495个植物病毒主题，并确定了数百个在结构蛋白和酶中发现的与基因调控无关的转录调控域。此外，该研究团队将ENTRAP-seq与机器引导设计结合起来，以半民族的方式设计植物转录因子的活性。他们的发现证明了在植物中部署可扩展的蛋白质功能分析将如何使植物中天然和合成编码多样性的表征成为可能。ENTRAP-seq系统地将蛋白质序列与原生植物环境中的转录输出联系起来。

研究人员表示，转录调控因子在植物生长发育和环境响应中发挥关键作用；然而，了解它们在蛋白质水平上的调控活性是如何被编码的，一直受到缺乏多通道大规模方法来表征植物蛋白质文库的阻碍。

附：英文原文

Title: Multiplexed profiling of transcriptional regulators in plant cells

Author: Alamos, Simon, Waldburger, Lucas, Dee, Amanda, Owens, Lauren A., Rattan, Rohan, Ong, Shirlyne, Shih, Patrick M.

Issue&Volume: 2025-11-25

Abstract: Transcriptional regulators play key roles in plant growth, development and environmental responses; however, understanding how their regulatory activity is encoded at the protein level has been hindered by a lack of multiplexed large-scale methods to characterize protein libraries in planta. Here we present enrichment of nuclear trans-elements reporter assay in plants with sequencing (ENTRAP-seq), a high-throughput method that introduces protein-coding libraries into plant cells to drive a nuclear magnetic sorting-based reporter, enabling multiplexed measurement of regulatory activity from thousands of protein variants. Using ENTRAP-seq and machine learning, we screen 1,495 plant viruses and identify hundreds of putative transcriptional regulatory domains found in structural proteins and enzymes not associated with gene regulation. In addition, we combine ENTRAP-seq with machine-guided design to engineer the activity of a plant transcription factor in a semirational fashion. Our findings demonstrate how scalable protein function assays deployed in planta will enable the characterization of natural and synthetic coding diversity in plants. ENTRAP-seq systematically links protein sequence to transcriptional output in a native plant context.

DOI: 10.1038/s41587-025-02880-w

Source: https://www.nature.com/articles/s41587-025-02880-w