PUBLICATION
Transcriptional Activity and DNA Methylation Dynamics of the Gal4/UAS System in Zebrafish
- Authors
- Pang, S.C., Wang, H.P., Zhu, Z.Y., Sun, Y.H.
- ID
- ZDB-PUB-150523-7
- Date
- 2015
- Source
- Marine biotechnology (New York, N.Y.) 17(5): 593-603 (Journal)
- Registered Authors
- Pang, Shaochen, Sun, Yonghua
- Keywords
- Gal4/UAS, Transcriptional activity, Transcriptional silencing, DNA methylation, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/metabolism
- DNA Methylation/genetics*
- Gene Expression Regulation, Developmental
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Zebrafish
- Zebrafish Proteins/genetics*
- PubMed
- 25997914 Full text @ Mar. Biotechnol.
Citation
Pang, S.C., Wang, H.P., Zhu, Z.Y., Sun, Y.H. (2015) Transcriptional Activity and DNA Methylation Dynamics of the Gal4/UAS System in Zebrafish. Marine biotechnology (New York, N.Y.). 17(5):593-603.
Abstract
The Gal4/upstream activating sequence (UAS) system is a powerful genetic tool for the temporal and spatial expression of target genes. In this study, the dynamic activity of the Gal4/UAS system was monitored in zebrafish throughout the entire lifespan and during germline transmission, using an optimized Gal4/UAS, KalTA4/4xUAS, which is driven by two muscle-specific regulatory sequences. We found that UAS-linked gene expression was transcriptionally amplified by Gal4/UAS during early developmental stages and that the amplification effects tended to weaken during later stages and even disappear in subsequent generations. In the F2 generation, the transcription of a UAS-linked enhanced green fluorescent protein (EGFP) reporter was transcriptionally silent from 16 days post-fertilization (dpf) into adulthood, yet offspring of this generation showed reactivation of the EGFP reporter in some strains. We further show that the transcriptional silencing and reactivation of UAS-driven EGFP correlated with the DNA methylation levels of the UAS regulatory sequences. Notably, asymmetric DNA methylation of the 4xUAS occurred in oocytes and sperm. Moreover, the paternal and maternal 4xUAS sequences underwent different DNA methylation dynamics after fertilization. Our study suggests that the Gal4/UAS system may represent a powerful tool for tracing the DNA methylation dynamics of paternal and maternal loci during zebrafish development and that UAS-specific DNA methylation should be seriously considered when the Gal4/UAS system is applied in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping