Community Action Needed: Please respond to the NIH RFI
ZFIN ID: ZDB-PUB-121004-9
Cautious use of fli1a:EGFP transgenic zebrafish in vascular research
Liu, Z., and Liu, F.
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.