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

项目摘要 细胞信号转导的研究从现有的改变信号转导的方法中获益良多。 以可控的方式描述调节细胞功能的特定分子事件的途径。 突变是一种用来移除特定识别位点或后识别位点的方法。 翻译修饰，以评估信号后果并推断 目标站点。虽然功能强大，但这种方法需要细胞发育、生长和激活才能 发生在非天然基因的背景下，这种基因可能具有辅助效应，使 实验结果。提交的申请书采用了一种全新的方法来研究 细胞信号转导；分子印迹纳米颗粒(MINP)是由严明开发的 对它们的短靶序列表现出精致的选择性和亲和力。细胞 渗透性已经被证明，在这一应用中，赵和安德烈奥蒂将测试 这些新试剂在T细胞信号转导研究中的有效性。具体的靶序列有 选择用于MINP生成的结合亲和力和特异性将在体外和细胞内进行测试 裂解产物，以及MINP在T细胞中的内化和靶向结合将得到充分的表征。如果 这些原理实验的成功，将为MINP的应用铺平道路 技术深入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.