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

基本信息

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
项目类别：
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

项目摘要

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 CHEETAH 中心成员提供制作、操作、并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）将提供使用不易获得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肽/蛋白质的组成，该方法非常适合生产含有标签（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成分）氨基酸、环状或交联肽/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质合成的范围扩大到更大、更具挑战性的目标。第 2 部分（蛋白质设计）将与 CHEETAH 中心实验室合作，使用和扩展尖端技术计算蛋白质设计方法，用于生成支持 HIV-1 的新型蛋白质工具、试剂和平台研究。具体进展将包括将深度学习方法纳入蛋白质设计流程中，并应用增强的设计方法来优化靶蛋白表达、稳定性和同质性。第 3 部分（CRISPR 筛选方法）将进行 CRISPR 验证实验和 HIV-CRISPR 筛选与现有的 CRISPR 文库合作，还开发新的文库来识别与表型相关的候选宿主基因猎豹中心实验室感兴趣。总体目标是：1）发现并突出功能相关的主机进一步研究的蛋白质目标，2) 为功能验证研究提供专业知识。