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Titolo:
Foreign DNA integration - Genome-wide perturbations of methylation and transcription in the recipient genomes
Autore:
Muller, K; Heller, H; Doerfler, W;
Indirizzi:
Univ Cologne, Inst Genet, D-50931 Cologne, Germany Univ Cologne Cologne Germany D-50931 nst Genet, D-50931 Cologne, Germany
Titolo Testata:
JOURNAL OF BIOLOGICAL CHEMISTRY
fascicolo: 17, volume: 276, anno: 2001,
pagine: 14271 - 14278
SICI:
0021-9258(20010427)276:17<14271:FDI-GP>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
DENOVO METHYLATION; MAMMALIAN GENOME; HAMSTER-CELLS; SEQUENCES; INSERTION; PATTERNS; ADENOVIRUS-TYPE-12; HYBRIDIZATION; FRAGMENTS; PROTOCOL;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
27
Recensione:
Indirizzi per estratti:
Indirizzo: Doerfler, W Univ Cologne, Inst Genet, Weyertal 121, D-50931 Cologne, Germany Univ Cologne Weyertal 121 Cologne Germany D-50931 ne, Germany
Citazione:
K. Muller et al., "Foreign DNA integration - Genome-wide perturbations of methylation and transcription in the recipient genomes", J BIOL CHEM, 276(17), 2001, pp. 14271-14278

Abstract

In hamster cells transgenic for the DNA of adenovirus type 12 (Ad12) or for the DNA of bacteriophage lambda, the patterns of DNA methylation in specific cellular genes or DNA segments remote from the site of transgene insertion were altered. In the present report, a wide scope of cellular DNA segments and genes was analyzed. The technique of methylation-sensitive representational difference analysis (MS-RDA) was based on a subtractive hybridization protocol after selecting against DNA segments that were heavily methylated and hence rarely cleaved by the methylation-sensitive endonuclease HpaII. The MS-RDA protocol led to the isolation of several cellular DNA segments that were indeed more heavily methylated in lambda DNA-transgenic hamstercell lines. By applying the suppressive subtractive hybridization technique to cDNA preparations from nontransgenic and Ad12-transformed or A DNA-transgenic hamster cells, several cellular genes with altered transcription patterns were cloned from Ad12-transformed or lambda DNA-transgenic hamster cells. Many of the DNA segments with altered methylation, which were isolated by a newly developed methylation-sensitive amplicon subtraction protocol,and cDNA fragments derived from genes with altered transcription patterns were identified by their nucleotide sequences. In control experiments, no differences in gene expression or DNA methylation patterns were detectable among individual nontransgenic BHK21 cell clones. In one mouse line transgenic for the DNA of bacteriophage lambda, hypermethylation was observed in the imprinted Igf2r gene in DNA from heart muscle. Two mouse lines transgenicfor an adenovirus promoter-indicator gene construct showed hypomethylationin the interleukin 10 and Igf2r loci. We conclude that the insertion of foreign DNA into an established mammalian genome can lead to alterations in cellular DNA methylation and transcription patterns. It is conceivable that the genes and DNA segments affected by these alterations depend on the site(s) of foreign DNA insertion.

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Documento generato il 25/11/20 alle ore 10:07:38