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 该核心旨在为猎豹中心成员提供制作，操纵， 并确定与我们中心正在研究的不同HIV-HOST系统的研究有关的新生物分子。 具体而言，我们用于生物聚合物合成和筛选核心的工具（核心1）将开发新的方法并提供 中心成员可以使用最先进的仪器和肽合成，蛋白质设计和专业知识 CRISPR筛选方法。 成分1（肽合成）将提供合成肽和蛋白质试剂，这些试剂不容易使用 传统的重组表达系统。由于化学合成提供了对 肽/蛋白质的组成，此方法非常适合生产含有标签的定制试剂（荧光团，， 同位素），修饰（甲基化，磷酸化）和非经典成分（镜像或其他不寻常的成分 氨基酸，环状或交联的肽/蛋白质）。此外，该核心将继续开发新型化学物质 肽合成工具将化学蛋白合成的覆盖范围扩展到更大且更具挑战性的靶标。 组件2（蛋白质设计）将与Cheetah Center Laboratories合作使用和扩展尖端 计算蛋白质设计方法，以生成新型蛋白质工具，试剂和平台以支持HIV-1 研究。具体的进步将包括将深度学习方法纳入蛋白质设计管道并应用 增强设计方法以优化靶蛋白表达，稳定性和同质性。 组件3（CRISPR筛选方法）将执行CRISPR验证实验和HIV-CRISPR屏幕 与现有的CRISPR库一起，还开发了新颖的图书馆来识别与表型有关的候选宿主基因 猎豹中心实验室感兴趣。总体目标是：1）发现并突出显示功能相关的主机 蛋白质目标进行进一步研究，2）为功能验证研究提供了专业知识。