Chemical approaches for precision genome editing

精确基因组编辑的化学方法

基本信息

批准号：
10378157
负责人：
Amit Choudhary
金额：
$ 35.26万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
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/10378157
关键词：
Acetyltransferase Affinity Base Pairing Binding Biomedical Technology CRISPR/Cas technology Cells Charge Chemicals Chromosomal translocation Clustered Regularly Interspaced Short Palindromic Repeats Complex Cysteine DNA DNA Binding Deacetylase Deoxyribonuclease I Development Dose Engineering Enzymes Esterification Gene Therapy Agent Genetic Transcription Genome Genome engineering Genomics Guide RNA HIV Hereditary Disease Hydrophobicity Image Impairment Laboratories Light Lysine Mammalian Cell Methods Modification Muscular Dystrophies Pathology Performance Permeability Proteins Proteome Rain Reagent Reporting Repression Ribonucleases Scheme Site Specificity Structure Structure-Activity Relationship System Technology Time Toxic effect Transcription Coactivator Transcription Repressor Transcriptional Activation Ursidae Family Vision Disorders base chemical genetics chemoproteomics endonuclease epigenome editing esterase gene repression gene therapy genetic approach genome editing genomic locus genotoxicity improved inhibitor mimetics nanomolar nuclease personalized approach programs small molecule small molecule inhibitor targeted nucleases

项目摘要

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