Viral Vector Core

病毒载体核心

基本信息

批准号：
10248197
负责人：
JERREL L YAKEL
金额：
$ 103.2万
依托单位：
NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
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/10248197
关键词：
2019-nCoV Adenoviruses Antiviral Agents Biochemical Biological Biological Assay Birds CRISPR/Cas technology Cattle Cell Line Cells Clinical Research Clustered Regularly Interspaced Short Palindromic Repeats Core Facility Coronavirus Developmental Biology Disease Epigenetic Process Family suidae Gene Delivery Gene Expression Gene Transfer Techniques Genes Genetic Genome Goals HIV Hematopoietic Hormone Receptor Human Resources Immune response Immunity Immunologics In Vitro Inflammation Influenza A virus Interphase Cell Intravenous Ion Channel Knock-out Laboratories Lentivirus Vector Lymphocytic choriomeningitis virus Methods Modification Mus National Institute of Environmental Health Sciences National Toxicology Program Neurobiology Neurons Primary Cell Cultures Production Protocols documentation Rabies Rattus Reagent Recombinants Reproductive Biology Research Research Personnel Research Support Respiratory syncytial virus Retroviridae Safety Sampling Services Signal Transduction Sindbis Virus Structure Study models Subfamily lentivirinae Supervision System Technology Time Tissues Toxicology Training Transgenic Animals Transgenic Organisms Validation Vesicular stomatitis Indiana virus Viral Viral Vector Virion Virus Zika Virus adult stem cell animal model development animal tissue blastomere structure cell immortalization cost design genome integrity human disease in vivo inducible gene expression knock-down protein expression protein purification small hairpin RNA stable cell line stem cell biology structural biology tool transcription factor vector viral gene delivery zygote

项目摘要

The Viral Vector Core facility currently offers support for production, purification, titering, and validation of twelve types of viruses: Adenovirus, Adenoassociated virus (AAV), coronaviruses (229E, OC43, and NL63), Influenza A, lentiviruses including human immunodeficiency virus (HIV), Lymphocytic choriomeningitis virus (LCMV), Rabies-delta-G (SAD-B19-dG, dGL, SiR, and CVS-N2c), Retrovirus, Respiratory syncytial virus (RSV), Sindbis Virus, vesicular stomatitis virus (VSV) and Zika virus. The core utilizes multiple vector systems to safely produce recombinant, pseudotyped and wild type viruses. This year, the Viral Vector Core also added services for propagation and transduction assays using SARS-CoV-2 pseudotyped virions and coronavirus strains 229E, OC43, and NL63. These transduction assays were used to screen a multitude of anti-viral reagents. SARS-CoV-2 pseudotyped virions were also utilized for structural studies of the virus spike. In addition, the core facility developed transduction protocols for gene delivery to primary or immortalized cell lines, mouse fertilized eggs, and animal tissue. In the past year, the Viral Vector Core has packaged and validated over 275 viral samples for gene delivery. These vectors have been utilized in research to express ion channels, transcription factors, biological indicators, hormone receptors, secreted protein expression and purification, gene knockdown by shRNA or genetic modification using CRISPR components. Viral vectors were also used to introduce genes into primary and immortalized cell lines, mouse and rat tissue, or cultured tissue. Other difficult to transfect cell lines, such as hematopoietic and mouse embryonic cells, were also successfully transduced using viral vectors. Lentiviral vectors facilitated immortalizing primary cell lines for toxicological studies and several viral strains were imported for use in immunological studies. The Viral Vector Core supports research in the Division of National Toxicology Program (DNTP), Clinical Research Branch (CRB), Epigenetic and Stem Cell Biology Laboratory (ESCBL), Genome Integrity and Structural Biology Laboratory (GISBL), Immunity, Inflammation and Disease Laboratory (IIDL), Neurobiology Laboratory (NL), Signal Transduction Laboratory (STL), and Reproductive and Developmental Biology Laboratory (RDBL)at the NIEHS by providing highly effective means of gene delivery.

病毒载体核心设施目前为 12 种病毒的生产、纯化、滴度测定和验证提供支持：腺病毒、腺相关病毒 (AAV)、冠状病毒（229E、OC43 和 NL63）、甲型流感、慢病毒（包括人类免疫缺陷病毒）（ HIV）、淋巴细胞脉络丛脑膜炎病毒（LCMV）、狂犬病-δ-G（SAD-B19-dG、 dGL、SiR 和 CVS-N2c)、逆转录病毒、呼吸道合胞病毒 (RSV)、辛德比斯病毒、水泡性口炎病毒 (VSV) 和寨卡病毒。该核心利用多个载体系统来安全地生产重组、假型和野生型病毒。今年，病毒载体核心还增加了使用 SARS-CoV-2 假型病毒粒子和冠状病毒株 229E、OC43 和 NL63 进行繁殖和转导测定的服务。这些转导测定用于筛选多种抗病毒试剂。 SARS-CoV-2 假型病毒颗粒也被用于病毒刺突的结构研究。此外，核心设施还开发了将基因传递至原代或永生化细胞系、小鼠受精卵和动物组织的转导方案。去年，病毒载体核心已包装并验证了超过 275 个用于基因传递的病毒样本。这些载体已用于表达离子通道、转录因子、生物指示剂、激素受体、分泌蛋白表达和纯化、通过 shRNA 进行基因敲除或使用 CRISPR 组件进行基因修饰的研究。病毒载体还用于将基因引入原代细胞系和永生化细胞系、小鼠和大鼠组织或培养组织中。其他难以转染的细胞系，如造血细胞和小鼠胚胎细胞，也可以使用病毒载体成功转导。慢病毒载体促进了原代细胞系的永生化以用于毒理学研究，并且进口了几种病毒株用于免疫学研究。病毒载体核心支持国家毒理学计划部门 (DNTP)、临床研究部门 (CRB)、表观遗传和干细胞生物学实验室 (ESCBL)、基因组完整性和结构生物学实验室 (GISBL)、免疫、炎症和疾病实验室的研究(IIDL)、神经生物学实验室 (NL)、信号转导实验室 (STL) 和 NIEHS 的生殖与发育生物学实验室 (RDBL)，通过提供高效的基因传递手段。