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

分子印迹纳米颗粒作为阐明 T 细胞信号传导事件的新工具

基本信息

批准号：
10452166
负责人：
AMY H ANDREOTTI
金额：
$ 22.95万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10452166
关键词：
Adaptor Signaling Protein Address Affinity Amino Acid Sequence Autoimmunity Binding Binding Sites Biological Assay CD3 Antigens CRISPR/Cas technology Cell membrane Cell physiology Cells Cellular biology Chemistry Collaborations Development Disease Elements Event Exhibits Family Generations Genes Goals Growth ITAM Immune Immune response Immunosuppression In Vitro Investigation Investments Knowledge LCP2 gene Masks Mediating Methods Micelles Molecular Molecular Target Morphologic artifacts Mutagenesis Mutation PXXP Motif Pathway interactions Permeability Phosphorylation Phosphorylation Site Phosphotransferases Phosphotyrosine Post-Translational Modification Site Post-Translational Protein Processing Preparation Prevalence Proline Proline-Rich Domain Proteins Proteome Protocols documentation Reagent Research Design Signal Pathway Signal Transduction Site Specificity Surface T-Lymphocyte Technology Testing Toxic effect Tyrosine Water base cell type combinatorial crosslink efficacy testing experimental study imprint in vitro testing knockout gene molecular site nanoparticle non-Native novel strategies novel therapeutics performance tests polyproline prevent protein aminoacid sequence protein complex protein protein interaction public health relevance receptor receptor function structural biology tool

项目摘要

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.

项目摘要细胞信号的研究已从现有方法中的改变信号传导中受益匪浅以受控方式描绘介导细胞功能的特定分子事件的途径。诱变是一种用于删除特定识别位点或后地点的方法翻译修改，评估信号后果并推断出的重要性目标站点。虽然强大，但这种方法需要细胞的开发，生长和激活才能发生在非本地基因的背景下，该基因可能具有辅助作用来混淆实验结果。提交的申请采取了一种全新的研究细胞信号传导；分子印迹纳米颗粒（MINPS）是由YAN开发的 Zhao具有出色的选择性和对其短目标序列的亲和力。细胞已经证明了渗透性，在此应用中，Zhao和Andreotti将测试这些新试剂在T细胞信号转导研究中的功效。特定目标序列是选择用于MINP的生成，结合亲和力和特异性将在体外和细胞中进行测试裂解物以及MINPS在T细胞中的内在化和目标结合将得到充分表征。如果成功，这些主要实验证明将为MINP的应用铺平道路技术到电池信号场的每个角落。