Premium
Expression of an Xist promoter‐luciferase construct during spermatogenesis and in preimplantation embryos: Regulation by DNA methylation
Author(s) -
Goto Tetsuya,
Christians Elisabeth,
Monk Marilyn
Publication year - 1998
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199804)49:4<356::aid-mrd2>3.0.co;2-m
Subject(s) - xist , biology , dna methylation , x inactivation , transgene , microbiology and biotechnology , methylation , blastocyst , dosage compensation , gene , regulation of gene expression , gene expression , genetics , genomic imprinting , epigenetics , transgenesis , luciferase , x chromosome , promoter , reproductive technology , embryogenesis , transfection
Dosage compensation for X‐linked genes in mammals is accomplished by inactivating one of the two X chromosomes in females, a process involving a regulatory gene, Xist (X‐inactive specific transcript). Xist maps to the X‐inactivation centre and is expressed from the inactive X chromosome in female somatic cells and at the time of X inactivation during spermatogenesis in the male. In female preimplantation embryos, Xist demonstrates imprinting in that the paternal allele inherited from the sperm is preferentially expressed. This preferential paternal Xist expression is correlated with paternal X inactivation in the extraembryonic lineages at the blastocyst stage. We have analysed a 233‐bp Xist promoter fragment (nt −220 to +13) for its ability to direct appropriate expression and its regulation by DNA methylation. This minimal promoter sequence directs expression of the luciferase reporter gene following injection of the construct into one‐cell embryos. In vitro methylation of the construct before injection represses transcription. In six different transgenic lines, expression of the Xist promoter‐luciferase transgene occurs only in the testis of the males (as for the endogenous Xist gene). The testis‐specific expression is correlated with hypomethylation of the transgene, although to different extents in different lines. Following paternal transmission, expression of the Xist promoter‐luciferase construct in preimplantation embryos is correlated with degree of hypomethylation in the testis and the degree of hypomethylation of the transgene in embryos at the morula stage. It is concluded that the patterns of methylation of the transgene in sperm (and in microinjected transgenes) can regulate the activity of the Xist promoter in the preimplantation embryo and thus support the hypothesis that gametic methylation patterns govern imprinted expression of the endogenous Xist gene in development. Mol. Reprod. Dev. 49:356–367, 1998. © 1998 Wiley‐Liss, Inc.