category
bioRxiv
date
Mar 18, 2026
slug
status
Published
summary
创新点包括:1) 开发无需转基因表达的CRISPR系统,直接在植物细胞核中操作;2) 利用dCas9-gRNA复合物实现特定染色质区域的高效特异性捕获;3) 适用于多种基因组区域(包括端粒、低拷贝和单拷贝序列);4) 为后续分析染色质相关蛋白复合物、DNA互作和RNA提供平台。
tags
基因编辑
核酸蛋白工具酶
type
Post
📄 原文题目
GRASP: A PLANT TRANSFORMATION-INDEPENDENT CRISPR-BASED SYSTEM FOR AFFINITY PURIFICATION OF SPECIFIC CHROMATIN LOCI
🔗 原文链接
💡 AI 核心解读
创新点包括:1) 开发无需转基因表达的CRISPR系统,直接在植物细胞核中操作;2) 利用dCas9-gRNA复合物实现特定染色质区域的高效特异性捕获;3) 适用于多种基因组区域(包括端粒、低拷贝和单拷贝序列);4) 为后续分析染色质相关蛋白复合物、DNA互作和RNA提供平台。
📝 英文原版摘要
Chromatin organization regulates genome stability and gene expression by controlling DNA accessibility to transcription factors and regulatory complexes. DNA-protein interactions are commonly investigated using chromatin immunoprecipitation (ChIP), which relies on specific antibodies often involving technically demanding protocols. CRISPR-Cas technologies have enabled sequence-specific targeting of genomic loci using catalytically inactive Cas9 (dCas9), but most CRISPR-based chromatin capture approaches in plants require transient or stable transformation to express the CRISPR machinery, limiting their applicability across species, tissues and physiological contexts. Here, we present GRASP (Genomic Region Affinity Sequestration by CRISPR-Purification), a transformation-independent strategy for sequence-specific chromatin isolation operating directly on purified plant nuclei. In GRASP, dCas9-gRNA ribonucleoprotein complexes are used to capture predefined genomic regions from chromatin under native conditions, bypassing the need for transgene expression. Using grapevine and tomato as model systems, we demonstrate efficient and highly specific enrichment of target loci, including telomeric repeats as well as low-copy and single-copy genomic regions, with qPCR and NGS validation. These results establish GRASP as a robust and broadly applicable platform for locus-specific chromatin isolation in plants. Beyond sequence-specific DNA isolation, GRASP establishes a versatile platform for potential downstream analyses of locus-associated chromatin components, including protein complexes, distal DNA-DNA interactions and chromatin-associated RNAs, providing new opportunities to investigate regulatory architecture in plant genomes.
- 作者:NotionNext
- 链接:https://tangly1024.com/article/32848bd6-1f96-8151-873b-ccd942763803
- 声明:本文采用 CC BY-NC-SA 4.0 许可协议,转载请注明出处。
相关文章
