Molecularly imprinted nanoparticles as new tools to elucidate T cell signaling events

项目摘要 细胞信号传导的研究从现有的改变信号传导的方法中获益匪浅 以受控的方式调控信号通路，以描绘介导细胞功能的特定分子事件。 诱变是一种这样的方法，用于去除特定的识别位点，或后处理位点。 翻译修饰，以评估信号传导后果并推断翻译修饰的重要性 目标部位。虽然强大，但这种方法需要细胞发育，生长和激活， 发生在非天然基因的背景下，这可能会产生辅助作用， 试验结果提交的申请采用了一种全新的方法来研究 细胞信号传导; Yan开发了分子印迹纳米颗粒（MINPs）， Zhao的基因对它们的短靶序列表现出精确的选择性和亲和力。细胞 渗透性已经得到证明，在这个应用中，Zhao和Andreotti将测试 这些新试剂在T细胞信号传导研究中的功效。特异性靶序列是 选择用于MINP生成的抗体，将在体外和细胞中测试结合亲和力和特异性。 将充分表征T细胞中MINPs的裂解物、内化和靶结合。如果 这些原理性实验的成功将为MINP的应用铺平道路 技术到细胞信号领域的每一个角落。

Project Summary The study of cellular signaling has benefitted greatly from existing approaches to alter signaling pathways in a controlled manner to delineate specific molecular events that mediate cell function. Mutagenesis is one such method used to remove a specific recognition site, or site of post- translational modification, to evaluate signaling consequences and deduce the importance of the target site. While powerful, this approach requires cell development, growth, and activation to take place in the context of a non-native gene which can have ancillary effects confounding the experimental results. The submitted application takes a completely new approach to the study of cellular signaling; molecularly imprinted nanoparticles (MINPs) have been developed by Yan Zhao that exhibit exquisite selectivity and affinity for their short target sequences. Cell permeability has been demonstrated and in this application Zhao and Andreotti will test the efficacy of these new reagents in the study of T cell signaling. Specific target sequences are chosen for MINP generation, binding affinity and specificity will be tested in vitro and in cell lysates, and internalization and target binding of MINPs in T cells will be fully characterized. If successful, these proof of principle experiments will pave the way for application of MINP technology to every corner of the cell signaling field.