HIV-1 host cell factors: a saturation screen for human chromosomes 4, 6, 21 and X

HIV-1 宿主细胞因子：人类 4、6、21 和 X 染色体的饱和筛选

• 批准号: 8011411
• 负责人: Nikunj V Somia
• 金额: $ 18.13万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8011411
• 关键词: Acquired Immunodeficiency Syndrome; Antiviral Agents; Bacillus amyloliquefaciens ribonuclease; Cell Communication; Cell Culture Techniques; Cell Line; Cells; Chromosomes, Human, Pair 4; Clone Cells; Code; Communicable Diseases; Complementary DNA; DNA; DNA Sequence; Drug Delivery Systems; Epidemic; Excision; Exons; Fluorescence; Future; Gene Expression; Genes; Genetic; Genetic Vectors; Genomics; Goals; HIV; HIV-1; Hand; Human; Human Cell Line; Human Chromosomes; Human Genome; Hypoxanthines; Infection; Insertional Mutagenesis; Introns; Knock-out; Life Cycle Stages; Link; Location; Mammalian Cell; Maps; Moloney Leukemia Virus; Monosomy; Mutagens; Mutation; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phenotype; Pilot Projects; Plasmids; Population; Process; Production; Promoter Regions; Proteins; RNA Splicing; Refractory; Replication Origin; Resistance; Resistance to infection; Resources; Sampling; Site; Sleeping Beauty; Structure-Activity Relationship; System; Technology; Testing; Transfection; Transferase Gene; Viral; Viral Drug Resistance; Viral Proteins; Viral Vector; Virus; base; combat; design; drug development; gene function; genome sequencing; interest; mutant; novel; novel strategies; preference; public health relevance; recombinase; small molecule; vector

DESCRIPTION (provided by applicant): HIV-1 host cell factors: a saturation screen for human chromosomes 4, 6, 21 and X. There is intense interest in identifying cellular host cell factors that modulate the life cycle of HIV-1. These factors can be classified as restriction factors that inhibit infection and permissive factors that aid the infection process. The study of these factors will aid in our understanding of viral and host cell interactions and to identify novel cellular drug targets. Since present drug therapies are based on targeting viral proteins, and the high mutation rate of HIV is able to generate resistance to these antiviral drugs, cellular proteins may constitute targets in which resistance is harder to generate. Various strategies to identify these proteins have been used. Here we propose an approach for the identification of host cell factors for the early life cycle of HIV. We aim to do a saturating insertional mutagenesis screen that will sample 1/5 of the human genome for host cell factor genes. We will achieve this aim using cells that have monosomies of chromosomes 4, 6, 21 and X. We will develop an insertional vector that disrupts gene function and select cells infected with this mutagenic vector for resistance to HIV-1 infection using an HIV-1 vector that is toxic on infection. Insertions into chromosome 4, 6, 21 and X will be identified through sequencing of DNA flanking the mutagenic vector. At the conclusion of this study we will have at least 2-3 verified host cell factors and the point in the lifecycle at which they are utilized. We will also have in hand a valuable resource - a knockout cell line of the host cell factor. This will enable structure-function relationships of the protein and the virus in future studies. These host cell factors may constitute novel targets for small drug molecules to combat HIV-1 and AIDS. PUBLIC HEALTH RELEVANCE: HIV-1 host cell factors: a saturation screen for human chromosomes 4, 6, 21 and X. HIV-1 relies on host cell proteins to complete its replication cycle. Identification of these factors is important since they are potential targets for the development of drugs to combat HIV-1 infection. In this study we aim to interrogate 1/5 the human genome in a pilot study to discover some host cell factors required by HIV-1 for infection.

描述（由申请人提供）： HIV-1 宿主细胞因子：人类 4、6、21 和 X 号染色体的饱和筛选。人们对鉴定调节 HIV-1 生命周期的细胞宿主细胞因子非常感兴趣。这些因素可分为抑制感染的限制因素和帮助感染过程的许可因素。对这些因素的研究将有助于我们了解病毒和宿主细胞的相互作用，并确定新的细胞药物靶点。由于目前的药物治疗是基于针对病毒蛋白，而HIV的高突变率能够对这些抗病毒药物产生耐药性，因此细胞蛋白可能成为较难产生耐药性的靶点。已经使用了多种策略来鉴定这些蛋白质。在这里，我们提出了一种鉴定艾滋病毒早期生命周期宿主细胞因子的方法。我们的目标是进行饱和插入诱变筛选，对 1/5 的人类基因组进行采样以寻找宿主细胞因子基因。我们将利用具有 4、6、21 和 X 号染色体单体的细胞来实现这一目标。我们将开发一种插入载体，破坏基因功能，并使用对感染有毒的 HIV-1 载体选择感染该诱变载体的细胞，以抵抗 HIV-1 感染。 4、6、21 和 X 号染色体的插入将通过对诱变载体侧翼的 DNA 进行测序来鉴定。在这项研究结束时，我们将获得至少 2-3 个经过验证的宿主细胞因子以及它们在生命周期中的利用点。我们还将拥有宝贵的资源——宿主细胞因子的敲除细胞系。这将使蛋白质和病毒在未来的研究中建立结构-功能关系。这些宿主细胞因子可能构成小药物分子对抗 HIV-1 和 AIDS 的新靶标。 公共卫生相关性：HIV-1 宿主细胞因子：人类 4、6、21 和 X 号染色体的饱和筛选。HIV-1 依赖宿主细胞蛋白来完成其复制周期。识别这些因素非常重要，因为它们是开发抗 HIV-1 感染药物的潜在目标。在这项研究中，我们的目标是在一项试点研究中询问 1/5 的人类基因组，以发现 HIV-1 感染所需的一些宿主细胞因子。