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Molecular and Cellular Biology, August 2001, p. 5008-5017, Vol. 21, No. 15
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.15.5008-5017.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Investigation of Elements Sufficient To Imprint the Mouse Air Promoter

Frank Sleutels and Denise P. Barlow*

Department of Molecular Genetics, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands

Received 24 January 2001/Returned for modification 23 March 2001/Accepted 9 May 2001

Imprinted maternal-allele-specific expression of the mouse insulin-like growth-factor type 2 receptor (Igf2r) gene depends on a 3.7-kb element named region 2, located in the second intron of the gene. Region 2 carries a maternal-allele-specific methylation imprint and contains an imprinted CpG island promoter (Air) that expresses a noncoding antisense RNA from the paternal inherited allele only. Here, we use transgenes to test the minimal requirements for imprinting of Air and to test if the action of region 2 is restricted to Igf2r. Transgenes up to 9 kb with Air as a single promoter are expressed but not imprinted. When coupled to the Igf2r CpG island promoter on a 44-kb transgene, Air was imprinted in one of three lines. However, Air on a 4.6-kb fragment is also imprinted in 2 of 14 lines when inserted in an intron of an adenine phosphoribosyltransferase (Aprt) transgene, and in one line, the imprinted methylation and expression of Air have been transferred onto the Aprt CpG island promoter. These data suggest that a dual CpG island promoter setting may facilitate Air imprinting as a short transgene and also show that Air can transfer imprinting onto other genes. However, for reliable Air imprinting, elements are necessary that are located outside a 44-kb region spanning the Air-Igf2r promoters.

* Corresponding author. Present address: ÖAW, Institute of Molecular Biology, Billrothstrasse 11, A-5020 Salzburg, Austria. Phone: 43 662 63961 14. Fax: 43 662 63961 29. E-mail: dbarlow{at}imb.oeaw.ac.at.

Molecular and Cellular Biology, August 2001, p. 5008-5017, Vol. 21, No. 15
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.15.5008-5017.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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