美国斯坦福大学Lacramioara Bintu和Michael C. Bassik研究组合作的最新研究，利用高通量测序揭示和表征了人转录效应因子。这一研究成果在线发表在2020年12月15日的国际学术期刊《细胞》上。

研究人员研发了高通量检测方法HT-recruit，其将蛋白质文库招募到报告基因，并通过测序测量其转录作用。使用这种方法，研究人员可以测量数千个结构域的基因沉默和激活。研究揭示了KRAB结构域抑制功能与进化年龄之间的关系，发现Homeodomain抑制因子的强度与Hox遗传组织共线，并确定了功能未知的多个结构域活性。

利用对CRISPRi KRAB的深度突变筛选，研究人员描绘了共抑制因子结合表面图谱，并鉴定出可提高稳定性/沉默的代替物。通过检测238种蛋白质，研究人员发现了短至十个氨基酸的抑制因子。

最后，研究鉴定了新的激活因子结构域，包括不同的KRAB。这些结果提供了包含效应因子等600种人类蛋白质的资源库，并揭示了探究蛋白质结构域功能的可扩展策略。

研究人员表示，成千上万的蛋白质定位于细胞核；然而，尚不清楚哪些蛋白包含转录效应因子。

附：英文原文

Title: High-Throughput Discovery and Characterization of Human Transcriptional Effectors

Author: Josh Tycko, Nicole DelRosso, Gaelen T. Hess, Aradhana, Abhimanyu Banerjee, Aditya Mukund, Mike V. Van, Braeden K. Ego, David Yao, Kaitlyn Spees, Peter Suzuki, Georgi K. Marinov, Anshul Kundaje, Michael C. Bassik, Lacramioara Bintu

Issue&Volume: 2020-12-15

Abstract: Thousands of proteins localize to the nucleus; however, it remains unclear which containtranscriptional effectors. Here, we develop HT-recruit, a pooled assay where proteinlibraries are recruited to a reporter, and their transcriptional effects are measuredby sequencing. Using this approach, we measure gene silencing and activation for thousandsof domains. We find a relationship between repressor function and evolutionary agefor the KRAB domains, discover that Homeodomain repressor strength is collinear withHox genetic organization, and identify activities for several domains of unknown function.Deep mutational scanning of the CRISPRi KRAB maps the co-repressor binding surfaceand identifies substitutions that improve stability/silencing. By tiling 238 proteins,we find repressors as short as ten amino acids. Finally, we report new activator domains,including a divergent KRAB. These results provide a resource of 600 human proteinscontaining effectors and demonstrate a scalable strategy for assigning functions toprotein domains.

DOI: 10.1016/j.cell.2020.11.024

Source: https://www.cell.com/cell/fulltext/S0092-8674(20)31541-5