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Chicken β‐globin insulator overcomes variegation of transgenes in Xenopus embryos
Author(s) -
Sekkali Belaïd,
Tran Hong Thi,
Crabbe Ellen,
De Beule Christophe,
Van Roy Frans,
Vleminckx Kris
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.07-098111
Subject(s) - xenopus , biology , transgene , chromatin , genetics , gene , gene silencing , insulator (electricity) , microbiology and biotechnology , locus control region , ctcf , genomic dna , scaffold/matrix attachment region , gene expression , promoter , chromatin remodeling , enhancer , electrical engineering , engineering
Chromatin structure and gene transcription regulation are intimately linked, and mosaic expression of randomly integrated transgenes into the genome is frequently observed. This variegation of transgene expression is likely due to the genomic integration site, which can affect the behavior of the integrated DNA sequence in a positive or a negative way. Insulators are a class of DNA elements that can protect genes from inappropriate signals emanating from their environment by acting as boundaries that prevent the spreading of nearby condensed chromatin that may otherwise silence expression. Here we show that transgenes escape this silencing in Xenopus laevis and Xenopus tropicalis embryos and that a stable, uniform, and heritable expression pattern is obtained when transgenes are flanked with tandem copies of the chicken P‐globin 5†HS4 insulator. Our data also indicate that the insulator confers copy‐number‐dependent transgene expression and can increase transgene expression from weak regulatory elements. Hence, it will be an invaluable tool for generating stable lines ex pressing different levels of a particular coding sequence.—Sekkali, B., Tran, H. T., Crabbe, E., De Beule, C., Van Roy, F., Vleminckx, K. Chicken β‐globin insulator overcomes variegation of transgenes in Xenopus embryos. FASEB J. 22, 2534–2540 (2008)