Mechanobiology Institute, National University of Singapore, Singapore.
Department of Nephrology, Hypertension and Rheumatology, University Hospital Münster, Münster, Germany.
EMBO Rep. 2022 Sep 5;23(9):e54401. doi: 10.15252/embr.202154401. Epub 2022 Jul 25.
YAP, an effector of the Hippo signalling pathway, promotes organ growth and regeneration. Prolonged YAP activation results in uncontrolled proliferation and cancer. Therefore, exogenous regulation of YAP activity has potential translational applications. We present a versatile optogenetic construct (optoYAP) for manipulating YAP localisation, and consequently its activity and function. We attach a LOV2 domain that photocages a nuclear localisation signal (NLS) to the N-terminus of YAP. In 488 nm light, the LOV2 domain unfolds, exposing the NLS, which shuttles optoYAP into the nucleus. Nuclear import of optoYAP is reversible and tuneable by light intensity. In cell culture, activated optoYAP promotes YAP target gene expression and cell proliferation. Similarly, optofYap can be used in zebrafish embryos to modulate target genes. We demonstrate that optoYAP can override a cell's response to substrate stiffness to generate anchorage-independent growth. OptoYAP is functional in both cell culture and in vivo, providing a powerful tool to address basic research questions and therapeutic applications in regeneration and disease.
YAP 是 Hippo 信号通路的效应因子，促进器官生长和再生。YAP 的持续激活会导致不受控制的增殖和癌症。因此，YAP 活性的外源性调节具有潜在的转化应用。我们提出了一种多功能的光遗传学构建体（optoYAP），用于操纵 YAP 的定位，从而调节其活性和功能。我们将 LOV2 结构域连接到 YAP 的 N 端，该 LOV2 结构域可光解核定位信号（NLS）。在 488nm 光下，LOV2 结构域展开，暴露出 NLS，将 optoYAP 转运到细胞核中。optoYAP 的核输入是可逆的，可以通过光强度进行调节。在细胞培养中，激活的 optoYAP 促进 YAP 靶基因的表达和细胞增殖。同样，optoYap 可用于斑马鱼胚胎中调节靶基因。我们证明 optoYAP 可以覆盖细胞对基质硬度的反应，从而产生非锚定依赖性生长。optoYAP 在细胞培养和体内都具有功能，为解决基础研究问题和再生与疾病的治疗应用提供了强大的工具。
