Myriad physiological and pathogenic processes are governed by protein levels and modifications. Controlled protein activity perturbation is essential to studying protein function in cells and animals. Based on Trim-Away technology, we screened for truncation variants of E3 ubiquitinase Trim21 with elevated efficiency (ΔTrim21) and developed multiple ΔTrim21-based targeted protein-degradation systems (ΔTrim-TPD) that can be transfected into host cells. Three ΔTrim-TPD variants are developed to enable chemical and light-triggered programmable activation of TPD in cells and animals. Specifically, we used ΔTrim-TPD for (1) red-light-triggered inhibition of HSV-1 virus proliferation by degrading the packaging protein gD, (2) for chemical-triggered control of the activity of Cas9/dCas9 protein for gene editing, and (3) for blue-light-triggered degradation of two tumor-associated proteins for spatiotemporal inhibition of melanoma tumor growth in mice. Our study demonstrates that multiple ΔTrim21-based controllable TPD systems provide powerful tools for basic biology research and highlight their potential biomedical applications.

无数的生理和致病过程受蛋白质水平和修饰的控制。受控的蛋白质活性扰动对于研究细胞和动物中的蛋白质功能至关重要。基于Trim-Away技术，我们筛选了效率更高的E3泛素酶Trim21的截短变体（ΔTrim21），并开发了多个可转染宿主细胞的基于ΔTrim21的靶向蛋白降解系统（ΔTrim-TPD）。开发了三种 ΔTrim-TPD 变体，以实现细胞和动物中 TPD 的化学和光触发可编程激活。具体来说，我们使用 ΔTrim-TPD（1）通过降解包装蛋白 gD 红光触发抑制 HSV-1 病毒增殖，（2）化学触发控制 Cas9/dCas9 蛋白的活性以进行基因编辑， (3)蓝光触发两种肿瘤相关蛋白的降解，从而时空抑制小鼠黑色素瘤的生长。我们的研究表明，多个基于 ΔTrim21 的可控 TPD 系统为基础生物学研究提供了强大的工具，并突出了它们潜在的生物医学应用。