Defining the functional boundaries of the Gata2 locus by rescue with a linked bacterial artificial chromosome transgene.

The Journal of Biological Chemistry
William BrandtJames Douglas Engel

Abstract

Transcription factor GATA-2 is vital for both hematopoietic progenitor cell function and urogenital patterning. Transgenic mapping studies have shown that the hematopoietic and urogenital enhancers are located hundreds of kbp 5' and 3' to the Gata2 structural gene, and both are vital for embryonic development. Because the size of mammalian genes, including all of their associated regulatory elements, can exceed a megabase, transgenic complementation in mice has, in specific instances, proven to be a formidable hurdle. After incorporating the Gata2 structural gene as well as the distant hematopoietic and urogenital enhancers into a single, contiguous piece of DNA by fusing two bacterial artificial chromosomes (BACs) into one, we formally tested the hypothesis that the functional boundaries of this locus are contained within this contiguous genomic span. We show that two independent lines of transgenic mice bearing a multicopy 413-kbp-linked Gata2 BAC transgene (bearing sequences from -187 to +226 kbp of the locus) are able to fully rescue Gata2 null mutant embryonic lethality and that the rescued animals behave and reproduce normally. Surprisingly, the linked BAC confers expression in the ureteric epithelium, whereas sequences w...Continue Reading

References

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Nov 16, 2004·Molecular and Cellular Biology·Melin KhandekarJames Douglas Engel
Feb 8, 2006·Proceedings of the National Academy of Sciences of the United States of America·Norio SuzukiMasayuki Yamamoto
Mar 31, 2007·Development·Melin KhandekarJames Douglas Engel

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Citations

Jun 4, 2013·Nature Communications·Shun YamazakiMasayuki Yamamoto
Sep 22, 2012·The Journal of Clinical Investigation·Kim-Chew LimJames Douglas Engel
Sep 22, 2012·The Journal of Clinical Investigation·Kirby D JohnsonEmery H Bresnick
Dec 31, 2015·Molecular Biology and Evolution·Alisha K HollowayKatherine S Pollard
Apr 18, 2009·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Daisuke NozawaMasayuki Yamamoto
Feb 25, 2015·British Journal of Haematology·Matthew CollinVenetia Bigley
Jan 25, 2014·The Journal of Biological Chemistry·Mark E Ebel, Geoffrey S Kansas
Dec 11, 2019·IUBMB Life·Mikiko SuzukiMasayuki Yamamoto

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