Career and Mentoring Development Program

生物的化学变化发生在细胞膜内部的隔间环境中。 系统地研究这种区隔的化学并利用它的好处进行治疗 通过定向酶进化的应用将需要控制合成和功能的方法 细胞样隔室的筛选。有指导的研究活动在#年的当前努力基础上显著扩大 通过探索可控高通量合成电路的微流控定向进化 用于体外隔室(IVC)的油中单分散水滴。这一战略正在实现新的 通过禁止单一优势基因型来探索RNA的催化适合度格局 在选择性扩增反应中占优势，夸大了群体的中性漂移。喷嘴 为这些实验开发了阵列微流控下腔静脉(MIVC)电路并实现了选择 包括L，每小时8个人。具有复杂表型的蛋白质的定向进化(运输， 膜显示、催化)将成为独立相研究的主题。PIVC系统 将用于合成单分散脂囊泡，用于隔室和功能展示 完整的膜蛋白、β-半乳糖苷酶和溶血素将作为使用pIVC的模型 处理器将在胞质和跨膜上进化出新的催化和选择性转运功能 分别为蛋白质。Long-Tennn研究计划的目标包括进化的膜受体(CCR5和 在脂泡中，选择增强病毒蛋白-受体复合体的结合，进化 结构-功能研究和膜结合可进化配体的合成及其在靶向研究中的应用 和诱饵疗法。

The chemical transformations of biology occur in a compartmentalized context inside the cell membrane. Systematically studying this compartmentalized chemistry and harnessing its benefits for therapeutic applications through directed enzyme evolution will require methods for controlled synthesis and functional screening of cell-like compartments. Mentored research activities significantly expanded on current efforts in microfluidic directed evolution by exploring circuitry for the controlled high-throughput synthesis of monodisperse water droplets in oil for in vitro compartmentalization (IVC). This strategy Is enabling new explorations of RNA's catalytic fitness landscape by prohibiting a single advantageous genotype from dominating in the selective amplification reaction, and exaggerating neutral drift ofthe population. A nozzle array microfluidic IVC (MIVC) circuit was developed for these experiments and enabled selections encompassing l e 8 individuals per hour. Directed evolution of proteins with complex phenotypes (transport, membrane display, catalysis) will form the theme for independent phase investigations. The pIVC system will be used to synthesize monodisperse lipid vesicles for compartmentalization and functional display of integral membrane proteins, P-galactosidase and hemolysin will serve as models for using the pIVC processor to evolve new catalytic and selective transport functions on cytosolic and transmembrane proteins, respectively. Long-tennn research program goals include evolving membrane receptors (CCR5 and CD4) in lipid vesicles, selecting for enhanced binding of viral protein-receptor complexes, evolutionary structure-function studies, and synthesizing membrane-bound evolvable ligands for applications In targeted and decoy therapeutics.