PUBLICATION
Testing of Cis-Regulatory Elements by Targeted Transgene Integration in Zebrafish Using PhiC31 Integrase
- Authors
- Hadzhiev, Y., Miguel-Escalada, I., Balciunas, D., Müller, F.
- ID
- ZDB-PUB-160729-24
- Date
- 2016
- Source
- Methods in molecular biology (Clifton, N.J.) 1451: 81-91 (Chapter)
- Registered Authors
- Balciunas, Darius, Hadzhiev, Yavor, Müller, Ferenc
- Keywords
- Microinjection, PhiC31 integrase, Transgenesis, Zebrafish
- MeSH Terms
-
- Animals
- Integrases/genetics
- Integrases/metabolism*
- Regulatory Sequences, Nucleic Acid/genetics*
- Transgenes/genetics*
- Zebrafish/genetics
- PubMed
- 27464802 Full text @ Meth. Mol. Biol.
Citation
Hadzhiev, Y., Miguel-Escalada, I., Balciunas, D., Müller, F. (2016) Testing of Cis-Regulatory Elements by Targeted Transgene Integration in Zebrafish Using PhiC31 Integrase. Methods in molecular biology (Clifton, N.J.). 1451:81-91.
Abstract
Herein we present several strategies for testing the function of cis-regulatory elements using the PhiC31 integrase system. Firstly, we present two different strategies to analyze the activity of candidate enhancer elements. Targeted integration of candidate enhancers into the same genomic location circumvents the variability-associated random integration and position effects. This method is suitable for testing of candidate enhancers identified through computational or other analyses a priori. Secondly, we present methodology for targeted integration of BACs into the same genomic location(s). By using additional reporters integrated into a BAC, this enables experimental testing whether cis-regulatory elements are functional in the sequence inserted in the BAC.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping