CHEETAH Center for the Structural Biology of HIV Infection, Restriction, and Viral Dynamics

CHEETAH HIV 感染、限制和病毒动力学结构生物学中心

• 批准号: 10663353
• 负责人: Michael S Kay
• 金额: $ 28.21万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-11 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663353
• 关键词: Amino Acids; Biochemical; Biology of HIV Infection; Biopolymers; CRISPR library; CRISPR screen; CRISPR/Cas technology; Cell Line; Cells; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Custom; Development; Epitopes; Genes; Genomic approach; Goals; HIV; HIV-1; Hand; Image; Infection; Integration Host Factors; Investigation; Isotopes; Label; Laboratories; Length; Lentivirus Vector; Libraries; Ligation; Link; Methodology; Methods; Methylation; Modification; Mutation; Peptide Synthesis; Peptides; Periodicity; Phase; Phenotype; Phosphorylation; Protein Engineering; Proteins; Reagent; Recombinants; Research; Role; SETDB1 gene; Solid; Solubility; System; TRIM Gene; Technology; Validation; Viral; base editing; candidate identification; chemical synthesis; crosslink; deep learning; design; experimental study; fluorophore; functional genomics; improved; insight; instrumentation; interest; learning strategy; machine learning model; member; method development; mutation screening; novel; novel strategies; peptide chemical synthesis; polypeptide; protein expression; protein structure; screening; structural biology; synthetic peptide; synthetic protein; tool; validation studies

CORE SUMMARY This Core is designed to provide CHEETAH Center members with access to novel approaches for making, manipulating, and identifying new biomolecules relevant to studies of different HIV-host systems under investigation in our Center. Specifically, our Tools for Biopolymer Synthesis and Screening Core (Core 1) will develop new methodology and provide Center members with access to state-of-the-art instrumentation and expertise in Peptide Synthesis, Protein Design, and CRISPR Screening Methodology. Component 1 (Peptide Synthesis) will provide synthetic peptide and protein reagents that are not readily accessible using traditional recombinant expression systems. Since chemical synthesis provides complete atomic control over the composition of peptides/proteins, this method is ideal for producing custom reagents containing labels (fluorophores, isotopes), modifications (methylation, phosphorylation), and non-canonical components (mirror-image or other unusual amino acids, cyclic or crosslinked peptides/proteins). Additionally, this Core will continue to develop novel chemical peptide synthesis tools to expand the reach of chemical protein synthesis to larger and more challenging targets. Component 2 (Protein Design) will collaborate with CHEETAH Center laboratories to use and extend cutting-edge computational protein design methodologies to generate novel protein tools, reagents, and platforms in support of HIV-1 research. Specific advances will include incorporating deep learning methods into protein design pipelines and applying the enhanced design approaches to optimize target protein expression, stability, and homogeneity. Component 3 (CRISPR Screening Methodology) will perform CRISPR validation experiments and HIV-CRISPR screens with existing CRISPR libraries, and also develop novel libraries to identify candidate host genes implicated in phenotypes of interest to CHEETAH Center laboratories. The overall goals are to: 1) uncover and highlight functionally relevant host protein targets for further study, and 2) provide expertise for functional validation studies.

核心摘要 此核心旨在为猎豹中心成员提供访问新方法的途径，以制作、操作 以及确定与我们中心正在调查的不同艾滋病毒宿主系统研究相关的新生物分子。 具体地说，我们的生物聚合物合成和筛选工具核心(核心1)将开发新的方法并提供 中心成员可以接触到最先进的仪器和多肽合成、蛋白质设计和 CRISPR筛查方法学。 组件1(多肽合成)将提供不容易使用的合成肽和蛋白质试剂 传统的重组表达系统。由于化学合成提供了对原子的完全控制 多肽/蛋白质的组合物，这种方法是生产包含标记(荧光团， 同位素)、修饰(甲基化、磷酸化)和非规范成分(镜像或其他不寻常的 氨基酸、环状或交联肽/蛋白质)。此外，这一核心将继续开发新的化学品 多肽合成工具，将化学蛋白质合成的范围扩大到更大和更具挑战性的目标。 组件2(蛋白质设计)将与猎豹中心实验室合作，使用和扩展尖端技术 用于产生支持HIV-1的新型蛋白质工具、试剂和平台的计算蛋白质设计方法 研究。具体进展将包括将深度学习方法纳入蛋白质设计流水线，并应用 优化目标蛋白表达、稳定性和同质性的增强设计方法。 构成部分3(CRISPR筛查方法学)将进行CRISPR验证实验和艾滋病毒-CRISPR筛查 与现有的CRISPR文库进行比较，并开发新的文库来鉴定与表型有关的候选宿主基因 猎豹中心实验室感兴趣的。总体目标是：1)发现并突出与功能相关的主持人 蛋白质靶标用于进一步研究，以及2)为功能验证研究提供专业知识。