Characterization of HIV Reservoirs in the Brain

大脑中 HIV 储存库的表征

• 批准号: 10678754
• 负责人: Avindra Nath
• 金额: $ 20万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 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/10678754
• 关键词: Address; Algorithms; Anti-Retroviral Agents; Autopsy; Bar Codes; Bioinformatics; Biological Assay; Brain; Cell Line; Consensus; DNA; DNA Library; DNA Probes; DNA sequencing; Detection; Environment; Equipment; Evolution; Genomic DNA; Genomics; Goals; HIV; HIV Genome; Human Genome; Infection; Length; Lymphoid; Lymphoid Tissue; Methods; Molecular; Patients; Penetration; Peripheral; Pharmaceutical Preparations; Preparation; Process; Proviruses; Site; Spleen; Techniques; Thalamic structure; Tissues; Tube; Variant; Viral; Viral Genes; Viral Proteins; Viral reservoir; base; design; digital; frontal lobe; genetic evolution; improved; integration site; nanopore; next generation sequencing; novel strategies; protein expression; sample collection; sequencing platform; single molecule; therapy design

We have just started sample collection and preparation for this project: 1) We have collected autopsy brain and peripheral lymphoid tissue from 4 patients with HIV. 2) We have synthesized digital droplet PCR primers for the detection of HIV reservoir in tissue. 3) We cultured 4 different cell lines for the extraction of genomic DNA and verification of integrated HIV genome with our synthesized primers. In addition, this genomic DNA will be used as positive control for our long read sequencing of HIV reservoir from HIV patient tissue. 4) We designed hybridization capture probe panel for the enrichment of HIV genomic fragment from the total genomic DNA extraction. 5) The equipment for this project is in the process of being purchased.

我们刚刚开始这个项目的样品采集和准备工作： 1)采集了4例HIV患者的尸检脑组织和外周淋巴组织。 2)合成了可用于检测组织中HIV储存库的数字滴状聚合酶链式反应引物。 3)培养了4个不同的细胞系，提取基因组DNA，并用合成的引物对整合的HIV基因组进行验证。此外，该基因组DNA将作为阳性对照，用于我们对HIV患者组织中HIV储存库的长读测序。 4)设计了杂交捕获探针板，用于从提取的总基因组DNA中提取HIV基因组片段。 5)本项目的设备正在采购中。