High throughput CRISPR-mediated functional validation of regulatory elements

高通量 CRISPR 介导的调控元件功能验证

• 批准号: 10240102
• 负责人: Bing Ren
• 金额: $ 157.19万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240102
• 关键词: Address; Biological Process; Biology; Candidate Disease Gene; Cell Line; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Sequence; Data; Disease; Elements; Gene Expression Regulation; Genes; Genetic Variation; Goals; Health; Human; Knowledge; Mammalian Cell; Maps; Mediating; Nucleic Acid Regulatory Sequences; Regulator Genes; Regulatory Element; Reporter; Tissues; Untranslated RNA; Validation; cell type; epigenomics; experimental study; genome editing; interest; mammalian genome; trait

Project Summary The overarching goal of the proposed study is to functionally characterize a large number of candidate functional elements in the mammalian genome. The ENCODE projects have revealed millions of putative regulatory elements across more than one hundred cell types and tissues. While these maps have significantly expanded our knowledge of non-coding sequences, there are still large gaps between having descriptive maps of functional elements and understanding the biology of these elements underlying gene regulation. These include: (a) few candidate functional elements predicted by the ENCODE experiments are functionally validated; (b) Epigenomic studies have not given/revealed information on the target genes of candidate functional elements. Therefore, it is still a challenge to interpret the biological functions of non-coding DNA sequences. To address these issues, the objective of this UM1 application is to perform large scale functional characterization of candidate functional elements in their native chromatin context. We will first identify candidate regulatory elements utilizing ENCODE data and generate reporter tagged genes of interest in cell lines utilizing a high throughput, automated platform. Second, we will interrogate candidate functional elements in their native chromatin contexts utilizing two complementary high throughput CRIPSR/Cas9 mediated genome editing approaches. We anticipate these analyses will significantly advance our knowledge of the biological functions of candidate regulatory regions and gene regulation in mammalian cells.

项目总结

项目成果

Parallel characterization of cis-regulatory elements for multiple genes using CRISPRpath.

• DOI: 10.1126/sciadv.abi4360
• 发表时间: 2021-09-17
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Ren X;Wang M;Li B;Jamieson K;Zheng L;Jones IR;Li B;Takagi MA;Lee J;Maliskova L;Tam TW;Yu M;Hu R;Lee L;Abnousi A;Li G;Li Y;Hu M;Ren B;Wang W;Shen Y
• 通讯作者: Shen Y