HIV-1 Genomic RNA Integrity

HIV-1 基因组 RNA 完整性

• 批准号: 10241263
• 负责人: Jeffrey M Kidd
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-18 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10241263
• 关键词: Applications Grants; Attenuated; Bioinformatics; Biological; Biological Assay; CRISPR interference; CRISPR library; CRISPR screen; Candidate Disease Gene; Cells; Complex; Enzymes; Exploratory/Developmental Grant; Genes; Genetic; Genetic Materials; Genetic Screening; Genome; Genomics; Goals; HIV Genome; HIV-1; High-Throughput Nucleotide Sequencing; Informatics; Lead; Lentivirus Vector; Libraries; Light; Logic; Mediating; Methodology; Methods; Molecular Profiling; Monitor; Northern Blotting; Nucleic Acids; Pathway interactions; Phenotype; Population; Preparation; Primates; Procedures; Property; Quality Control; RNA; RNA Processing; RNA Viruses; RNA-Directed DNA Polymerase; Records; Reproducibility; Research; Retroviridae; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; Running; Structure; Testing; Time; Virion; Virus; Virus Diseases; Virus Replication; Work; base; design; dimer; experience; experimental study; genome integrity; genomic RNA; innovative technologies; insight; mindfulness; monomer; mutant; novel; novel strategies; success; technology development; vector; viral genomics; whole genome

HIV-1 genomic RNA integrity Retroviruses are the only viruses that encapsidate two copies of their genetic material. One possible reason for this is that retroviral RNA appears to be damaged prior to reverse transcription; therefore, the presence of two copies of viral genomic RNA allows for successful reverse transcription through template switching. When virion RNA is examined on non-denaturing northern blots, it runs as a single dimer band. However, when heat- denatured to disrupt the dimer linkage, virion RNA includes a monomer band of reduced intensity compared to the dimer and also a smear of more rapidly-migrating RNAs, suggesting encapsidated RNA is extensively fragmented. Our central hypothesis is that this fragmentation of HIV-1 genomic RNA is due to the action of one or more host enzymes, the absence of which would lead to virions with more intact genomic RNAs and higher infectious titers. The objective of this proposal is to identify cellular genes that are associated with HIV- 1 RNA nicking using unbiased genetic screens and to test if these represent a new class of restriction factors. The proposed work combines Telesnitsky lab expertise in retroviral RNA with Tai lab expertise in whole- genome genetic screens for cellular factors that modulate viral infection plus Kidd lab expertise in specialized library preparation and bioinformatics, and consists of two aims: In Aim 1: we will seek to identify cellular genes that are required for observed damage to encapsidated HIV-1 RNAs using a whole-genome CRISPR library encoded in a lentiviral vector, and will validate candidate genes by using genetic depletion and rescue experiments. In Aim 2: we will assess properties of virion RNA fragmentation and examine its ramifications for viral replication. If successful, this project may reveal a new class of cellular restriction factors, may help illuminate novel cellular RNA processing pathways if the identified genes have not previously been shown to participate in RNA quality control, and may provide new insight into virion RNA structures and accessibility.

HIV-1基因组RNA完整性

项目成果

Directed evolution of potent neutralizing nanobodies against SARS-CoV-2 using CDR-swapping mutagenesis.

• DOI: 10.1016/j.chembiol.2021.05.019
• 发表时间: 2021-09-16
• 期刊: Cell chemical biology
• 影响因子: 8.6
• 作者: Zupancic JM;Desai AA;Schardt JS;Pornnoppadol G;Makowski EK;Smith MD;Kennedy AA;Garcia de Mattos Barbosa M;Cascalho M;Lanigan TM;Tai AW;Tessier PM
• 通讯作者: Tessier PM