PUBLICATION
The role of post-transcriptional RNA processing and plasmid vector sequences on transient transgene expression in zebrafish
- Authors
- Chatterjee, S., Min, L., Karuturi, R.K., and Lufkin, T.
- ID
- ZDB-PUB-090814-3
- Date
- 2010
- Source
- Transgenic Research 19(2): 299-304 (Journal)
- Registered Authors
- Keywords
- krt4 (keratin 4), EGFP (Enhanced Green Fluorescent Protein), Rabbit beta-globin intron, Microinjection, Transient assay, Biostatistics
- MeSH Terms
-
- Animals, Genetically Modified
- Transgenes/genetics
- Transgenes/physiology*
- beta-Globins/genetics
- Rabbits
- Base Sequence*
- RNA Processing, Post-Transcriptional*
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism*
- Introns/genetics
- Microinjections
- Animals
- Genetic Vectors/genetics*
- Plasmids/genetics*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Keratin-4/genetics
- Keratin-4/metabolism
- PubMed
- 19662507 Full text @ Transgenic. Res.
Citation
Chatterjee, S., Min, L., Karuturi, R.K., and Lufkin, T. (2010) The role of post-transcriptional RNA processing and plasmid vector sequences on transient transgene expression in zebrafish. Transgenic Research. 19(2):299-304.
Abstract
A tissue-specific transgenic model was employed to test the effects of intron and vector sequences on transgene expression in zebrafish after microinjection. In this model, the 2.3 kb promoter taken from the 5' upstream region of the transcription initiation site of keratin 4 (krt4) was used to drive the enhanced green fluorescence protein (EGFP) reporter gene in a transgenic vector. For assaying the strength of EGFP expression, the effects of including an intron before the EGFP coding region or using different forms of DNA, including circular plasmid, linear full-length plasmid, and the linear transgene coding region without any prokaryotic vector sequence, were tested. After microinjection, the transgene expression was analyzed using transient assays. Consequently, further comparative analysis supported by Fisher's exact test was performed based on the data generated by analyzing the strength of the transgene expression. It was shown that inclusion of an intron in the construct increases the transgene expression in a transient transgenic zebrafish assay. Furthermore, the circular plasmid containing the transgene produced the strongest EGFP expression.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping