Core 1: Viral Vector Core

核心1：病毒载体核心

• 批准号: 10415190
• 负责人: KRISTINE E YODER
• 金额: $ 14.92万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-21 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10415190
• 关键词: Binding; Biological Assay; CRISPR/Cas technology; Cancer Model; Cells; Cloning; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Consult; Coupled; Custom; DNA; Elementary Particles; Engineering; Enzyme-Linked Immunosorbent Assay; Feline Immunodeficiency Virus; Gene Expression; Genes; Genetic Engineering; Genetic Recombination; Genetic Transcription; Genome; Guide RNA; HIV-1; Human T-lymphotropic virus 1; Individual; Initiator Codon; Integrase; Laboratories; Leadership; Lentivirus Vector; Luciferases; Methodology; Methods; Modification; Murine leukemia virus; Mutation; Neomycin; Pathogenicity; Phosphorylation; Point Mutation; Proteomics; Proviruses; Puromycin; RNA Binding; RNA-Directed DNA Polymerase; Reagent; Reproducibility; Research Personnel; Resources; Response Elements; Retroviral Vector; Retroviridae; Services; Silent Mutation; Spumavirus; Taxes; Time; Transgenes; Variant; Vesicular stomatitis Indiana virus; Viral; Viral Genome; Viral Vector; Visual; Work; base; cell type; cost; delivery vehicle; design; expression vector; fusion gene; gene function; genetic manipulation; genome editing; homologous recombination; hygromycin A; interest; mutant; novel; overexpression; particle; prevent; promoter; prototype; repaired; selective expression; skills; small hairpin RNA; success; tool; vector; vector control

PROJECT SUMMARY – CORE 1 The Viral Vector Core (Core 1) will provide customized lentiviral or retroviral vectors to all PPG projects. Lentiviral or retroviral vector particles are fundamental reagents necessary for the efficient genetic engineering of HTLV-1 infected cells. The vector particles will be used to overexpress or reduce expression of host, viral, or novel fusion genes. Reduced expression will be achieved by delivery of either shRNA or CRISPR/Cas9. The vectors for reduced expression will be matched with vectors for complementation with wild type and functional mutant genes with silent mutations to prevent shRNA or CRISPR recognition. Lentiviral vector particles will also provide fluorescent and/or selection genes. Homology directed repair induced by a CRISPR/Cas9 vector will modify the HTLV-1 genome sequence. Integrase defective lentiviral vectors will be used to quantify the off- target effects of all CRISPR/Cas9 vectors. The Core will work with individual PPG labs to develop CRISPR/Cas9 reagents and methodologies for modification of HTLV-1 infected cells. The Core will also develop a novel vector that limits Cas9 expression both spatially and temporally to transcriptionally active HTLV-1 infected cells and reduce potential off-target editing. Specific aims of the core are: Aim 1 to provide custom retroviral vectors, Aim 2 to design, validate, and optimize CRISPR/Cas9 vectors, Aim 3 to develop an inducible, self-limiting HTLV-1 CRISPR/Cas9 vector. The lentiviral or retroviral vector particles produced by the Core will service Projects 1-3 of this PPG.

项目概要-核心1 病毒载体核心（核心1）将为所有PPG项目提供定制的慢病毒或逆转录病毒载体。 慢病毒或逆转录病毒载体颗粒是高效基因工程所必需的基本试剂 HTLV-1感染的细胞。载体颗粒将用于过表达或减少宿主、病毒或宿主细胞的表达。 新型融合基因减少的表达将通过递送shRNA或CRISPR/Cas9来实现。的 用于降低表达的载体将与用于与野生型和功能性互补的载体匹配 沉默突变的突变基因，以防止shRNA或CRISPR识别。慢病毒载体颗粒将 还提供荧光和/或选择基因。由CRISPR/Cas9载体诱导的同源定向修复 将改变HTLV-1基因组序列整合酶缺陷型慢病毒载体将用于定量脱- 所有CRISPR/Cas9载体的靶向效应。核心将与各个PPG实验室合作， 用于修饰HTLV-1感染的细胞的CRISPR/Cas9试剂和方法。核心也将 开发一种新的载体，在空间和时间上限制Cas9的表达， HTLV-1感染的细胞，并减少潜在的脱靶编辑。该核心的具体目标是： 定制逆转录病毒载体，目标2设计，验证和优化CRISPR/Cas9载体，目标3开发 可诱导的自限性HTLV-1 CRISPR/Cas9载体。由本发明的方法产生的慢病毒或逆转录病毒载体颗粒可用于治疗癌症。 核心将服务于本PPG的项目1-3。