Cautious use of fli1a:EGFP transgenic zebrafish in vascular research
- Authors
- Liu, Z., and Liu, F.
- ID
- ZDB-PUB-121004-9
- Date
- 2012
- Source
- Biochemical and Biophysical Research Communications 427(1): 223-226 (Journal)
- Registered Authors
- Liu, Feng, Liu, Zhibin
- Keywords
- transgenics, fli1a:EGFP, whole-mount in situ hybridization, blood vessel, zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/embryology*
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/metabolism
- Artifacts*
- Base Sequence
- Blood Vessels/embryology*
- Blood Vessels/metabolism
- Developmental Biology*
- Embryo, Nonmammalian/blood supply
- Embryo, Nonmammalian/metabolism
- Gene Expression Profiling
- Genetic Vectors
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism*
- In Situ Hybridization/standards*
- Molecular Sequence Data
- Plasmids/genetics
- Proto-Oncogene Protein c-fli-1/genetics
- Proto-Oncogene Protein c-fli-1/metabolism*
- RNA Probes/genetics
- Transgenes
- Zebrafish/embryology*
- Zebrafish/genetics
- Zebrafish/metabolism
- PubMed
- 22995292 Full text @ Biochem. Biophys. Res. Commun.
Integration of exogenous sequence into an intact genome may cause some artificial phenotype or unspecific observations. We noticed that there is unspecific vascular expression when using fli1a:EGFP transgenic embryos for whole-mount in situ hybridization (WISH) experiments. We therefore tested whether the residual vector sequence contained in the fli1a:EGFP transgene or the integration of transgene into the genome may cause this expression ‘noise’ and/or deregulation of gene expression at a genome-wide level. RNA probes were synthesized using two different methods, i.e. vector-based and PCR-based. The vector-based dnmt3 probe showed unspecific vascular expression in fli1a:EGFP embryos, but not in wildtype embryos, by WISH. Moreover, we also found that compared to that in wildtype, there were alterations in gene expression at whole-genome level in the fli1a:EGFP embryos. Our finding that the vector sequence contained in the fli1a:EGFP genome causes unspecific vascular expression by WISH and the genome-wide expression profiling is altered in fli1a:EGFP embryos strongly argue that extra caution should be taken for data interpretation when using transgenics, such as fli1a:EGFP, in developmental biology studies.