Career and Mentoring Development Program

职业和指导发展计划

• 批准号: 8077363
• 负责人: STEVEN Collins COLLINS
• 金额: $ 6.82万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8077363
• 关键词: Area; Binding; Biology; Breast; Catalysis; Catalytic RNA; Cell membrane; Cells; Chemicals; Chemistry; Chemokine (C-C Motif) Receptor 5; Clinical; Communities; Complex; Country; Educational workshop; Environment; Enzymes; Evolution; Exposure to; Funding; Future; Galactosidase; Genotype; Goals; Grant; Growth; Gynecologic; Hemolysin; Hour; In Vitro; Individual; Institution; Integral Membrane Protein; Investigation; Leadership; Learning; Ligands; Lipids; Lung; Malignant neoplasm of ovary; Malignant neoplasm of prostate; Membrane; Mentors; Methods; Microfluidics; Modeling; Nature; Oils; Organ; Ovarian; Ovary; Phase; Phenotype; Population; Program Development; Prostate; Proteins; Reaction; Recording of previous events; Reproduction spores; Research; Research Activity; Research Personnel; Schedule; Scientist; Screening procedure; Senior Scientist; Site; Specimen; Structure; System; Therapeutic; Translational Research; Transmembrane Transport; Vesicle; Viral Proteins; Water; Woman; Work; anticancer research; career; design; directed evolution; experience; fitness; gastrointestinal; interdisciplinary approach; interest; malignant breast neoplasm; meetings; programs; repository; research study; skills; success; symposium

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.

生物学的化学转化发生在细胞膜内的一个分隔环境中。 系统地研究这种划分的化学物质，并利用其对治疗的益处， 通过定向酶进化的应用将需要控制合成和功能性的方法， 细胞样区室的筛选。指导下的研究活动大大扩大了目前的努力， 微流控定向进化通过探索电路的控制高通量合成 油中的单分散水滴用于体外区室化（IVC）。这一战略使新的 探索RNA的催化适应性景观，禁止一个单一的优势基因型， 在选择性扩增反应中起主导作用，并夸大了群体的中性漂移。喷嘴 阵列微流控IVC（MIVC）电路被开发用于这些实验并使选择成为可能 包括每小时1 e 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）将形成独立阶段研究的主题。pIVC系统 将用于合成单分散脂质囊泡，用于区室化和功能展示， 整合膜蛋白、β-半乳糖苷酶和溶血素将作为使用pIVC的模型 在胞质和跨膜上进化新的催化和选择性转运功能的处理器 蛋白质，分别。长期研究计划的目标包括进化膜受体（CCR 5和 CD 4）在脂质囊泡中，选择增强病毒蛋白-受体复合物的结合，进化 结构-功能研究，并合成膜结合的可进化配体，用于靶向 和诱饵疗法