Development of a CRISPR-FISH toolset for imaging of genomic loci in living plants and its application to study the centromere structure and the process of chromosome condensation in living cells

开发用于活体植物基因组位点成像的 CRISPR-FISH 工具集及其在活细胞着丝粒结构和染色体浓缩过程研究中的应用

• 批准号: 358939560
• 负责人: Privatdozent Dr. Andreas Houben
• 金额: --
• 依托单位: Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/358939560?language=en
• 关键词: Development; CRISPR; FISH; toolset; imaging

Elucidating how the genome is spatio-temporally organised inside the nucleus is imperative to understand the regulation of genes and non-coding sequences during development. Emerging techniques of chromatin imaging promise to bridge the long-standing gap between sequencing studies disclosing genomic information and imaging studies providing spatial and temporal information. The project aims to harness the CRISPR-Cas9 system to develop a pioneering tool to visualize defined genomic loci in living cells of eudicot- (A. thaliana, carrot, N. benthamiana) and monocot (wheat) plants. This innovative technology will be used to analyse the centromere organization and the process of chromosome condensation in living plant cells.

阐明基因组是如何在细胞核内时空组织的，对于理解基因和非编码序列在发育过程中的调控是必不可少的。新兴的染色质成像技术有望弥合基因组信息测序研究和提供空间和时间信息的成像研究之间的长期差距。该项目旨在利用CRISPR-Cas9系统开发一种开创性的工具，以可视化真叶植物(A.thaliana、胡萝卜、N.benthamiana)和单子叶植物(小麦)活细胞中已定义的基因组位置。这项创新技术将用于分析植物活细胞的着丝粒组织和染色体凝聚过程。

项目成果

Application of Tris-HCl Allows the Specific Labeling of Regularly Prepared Chromosomes by CRISPR-FISH

Tris-HCl 的应用允许通过 CRISPR-FISH 对常规制备的染色体进行特异性标记

• DOI: 10.1159/000506720
• 发表时间: 2020
• 期刊: Cytogenetic and Genome Research
• 影响因子: 1.7
• 作者: Potlapalli BP;Schubert V;Metje-Sprink J;Liehr T;Houben A
• 通讯作者: Houben A

CRISPR/Cas9-Based RGEN-ISL Allows the Simultaneous and Specific Visualization of Proteins, DNA Repeats, and Sites of DNA Replication

• DOI: 10.1159/000502600
• 发表时间: 2019-11-01
• 期刊: CYTOGENETIC AND GENOME RESEARCH
• 影响因子: 1.7
• 作者: Nemeckova, Alzbeta;Waesch, Christina;Houben, Andreas
• 通讯作者: Houben, Andreas

Application of Aptamers Improves CRISPR-Based Live Imaging of Plant Telomeres

• DOI: 10.3389/fpls.2020.01254
• 发表时间: 2020-08-20
• 期刊: FRONTIERS IN PLANT SCIENCE
• 影响因子: 5.6
• 作者: Khosravi, Solmaz;Schindele, Patrick;Houben, Andreas
• 通讯作者: Houben, Andreas