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

核心摘要 这个核心旨在为切塔中心的成员提供新的方法来制作，操纵， 并确定新的生物分子相关的研究不同的艾滋病毒宿主系统正在调查中，在我们的中心。 具体来说，我们的生物聚合物合成和筛选核心工具（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 CRISPR筛选方法。 组件1（肽合成）将提供合成肽和蛋白质试剂，其不容易使用 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， 肽/蛋白质的组成，该方法对于生产含有标记物（荧光团， 同位素）、修饰（甲基化、磷酸化）和非典型组分（镜像或其他不寻常的 氨基酸、环状或交联肽/蛋白质）。此外，该核心将继续开发新的化学品 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CHEETAH中心实验室合作，使用和扩展尖端的 计算蛋白质设计方法，以生成支持HIV-1的新型蛋白质工具、试剂和平台 research.具体的进展将包括将深度学习方法纳入蛋白质设计管道，并将其应用于 增强的设计方法，以优化靶蛋白的表达、稳定性和均一性。 组件3（CRISPR筛选方法学）将进行CRISPR验证实验和HIV-CRISPR筛选 利用现有的CRISPR文库，并开发新的文库来鉴定与表型有关的候选宿主基因， CHEETAH中心实验室感兴趣。总体目标是：1）发现和突出功能相关的主机 蛋白质靶点，用于进一步研究; 2）为功能验证研究提供专业知识。

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.