使用CRISPR介导激活在基因组中的转录增强剂的表征
Jeong Hoon Han1, Hong Jo Lee2, Tae Hyun Kim1,3
1Department of Animal Science, The Pennsylvania State University, University Park, PA, United States.
Frontiers in genome editing
|November 13, 2023
概括
克里斯普尔激活 (CRISPRa) 精确地准肉DNA调节元件,以识别功能增强剂. 这种方法有效验证非编码区域,推进遗传理解和育种计划.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 基因规则 基因规则
背景情况:
- DNA调节元件控制基因表达,非编码区域的变异对于理解遗传特征至关重要.
- 识别功能性调节元素对于生物背景和在育种计划中的潜在应用至关重要.
- 克里斯普技术为研究非编码监管元素提供了一种强大的方法.
研究的目的:
- 确定与HBBA,IRF7和PPARG基因相关的基因组中的促进子和假定增强子区域.
- 使用CRISPR激活 (CRISPRa) 功能验证已识别的增强器区域.
- 通过准调控元素来证明CRISPRa在操纵内源基因表达方面的有效性.
主要方法:
- 从动物基因组功能注释项目中利用表观遗传数据.
- 设计的指导RNA向促进器和候选增强器区域.
- 在的DF-1细胞中利用dCas9-p300和dCas9-VPR的CRISPR激活 (CRISPRa).
主要成果:
- 通过比较促进剂激活后的基因表达与促进剂和增强剂的协同激活后识别的功能增强剂.
- 当功能增强剂与促进剂联合激活时,观察到目标基因的增强上调.
- 通过有针对性的调节元素,证明了内源基因表达的精确操纵.
结论:
- 克里斯普拉对基因组中非编码调控元素的功能验证非常有效.
- 这种方法精确地操纵内源基因表达,为繁殖计划的遗传改进提供了潜在的潜力.
- 这项研究强调了CRISPRa在剖析复杂的遗传调节网络方面的力量.
相关概念视频
RNA Polymerase II Accessory Proteins
9.2K
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
9.2K
Eukaryotic Transcription Activators
11.1K
Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
11.1K
Combinatorial Gene Control
8.4K
Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
8.4K
Co-activators and Co-repressors
7.4K
Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
7.4K


