期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 427, 期 1, 页码 223-226出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2012.09.064
关键词
Transgenics; fli1a:EGFP; Whole-mount in situ hybridization; Blood vessel; Zebrafish
资金
- National Basic Research Programs of China [2010CB945300, 2011CB943900]
- National Science Foundation of China [30971678, 31000647]
- Strategic Priority Research Program of the CAS [XDA01010110]
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. (C) 2012 Elsevier Inc. All rights reserved.
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