Chemical approaches for precision genome editing

精确基因组编辑的化学方法

• 批准号: 10389932
• 负责人: Amit Choudhary
• 金额: $ 7.4万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10389932
• 关键词: Acetyltransferase; Biomedical Technology; CRISPR/Cas technology; Chemicals; Chromosomal translocation; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Deacetylase; Enzymes; Gene Therapy Agent; Genetic Transcription; Genome; Genome engineering; Genomics; Guide RNA; HIV; Hereditary Disease; Image; Impairment; Methods; Muscular Dystrophies; Pathology; Reagent; Repression; Site; Specificity; Technology; Transcription Coactivator; Transcription Repressor; Transcriptional Activation; Vision Disorders; base; chemical genetics; endonuclease; epigenome editing; gene repression; genetic approach; genome editing; genomic locus; nuclease; personalized approach

PROJECT SUMMARY CRISPR-Cas9 is an RNA-guided endonucleases that is being actively used for sequence-specific DNA recognition, genome engineering, targeted transcriptional activation/repression and genome imaging. Cas9 is being developed as a gene therapy agent for multiple pathologies, including HIV, vision disorders, muscular dystrophy, and hereditary disorders. The precision control of specificity of CRISPR-Cas9 is required as off- target effects and chromosomal translocations are observed at elevated activity. Further, the ease of targeting catalytically impaired Cas9 to any genomic locus has resulted in transformative technologies. For example, the fusion of catalytically inactive Cas9 (dCas9) to transcriptional activators or repressors has enabled gene transcription and repression; fusion of catalytically impaired Cas9 to base-modifying enzymes has allowed base conversion (e.g., C→T) at specific genomic sites; dCas9‒GFP fusion has made imaging genomic loci possible; and dCas9‒acetyltransferases or deacetylases fusion has enabled epigenome editing. We propose to apply chemical and genetic approaches develop reagents and methods that will allow precision control of specificity of CRISPR-Cas9. 1

项目概要 CRISPR-Cas9 是一种 RNA 引导的核酸内切酶，被积极用于序列特异性 DNA 识别、基因组工程、靶向转录激活/抑制和基因组成像。 正在被开发为多种疾病的基因治疗剂，包括艾滋病毒、视力障碍、肌肉疾病 营养不良和遗传性疾病需要精确控制 CRISPR-Cas9 的特异性。 在活性升高时观察到靶向效应和染色体易位。此外，靶向的容易性。 任何基因组位点的催化受损 Cas9 都带来了变革性技术。 催化失活的 Cas9 (dCas9) 与转录激活子或阻遏子的融合使基因 转录和抑制；催化受损的 Cas9 与碱基修饰酶的融合允许碱基 特定基因组位点的转换（例如 C→T）；dCas9-GFP 融合使基因组位点成像成为可能； 我们建议应用dCas9-乙酰转移酶或脱乙酰酶融合来实现表观基因组编辑。 化学和遗传方法开发能够精确控制特异性的试剂和方法 CRISPR-Cas9。 1