Methods for assaying gene imprinting and methylated cpg islands

a technology of methylation and cpg islands, which is applied in the direction of genetically modified cells, drug compositions, skeletal/connective tissue cells, etc., can solve the problems of hampered experimental studies of the timing and mechanism of genomic imprinting
US20050153440A1Inactive Publication Date: 2005-07-14THE JOHN HOPKINS UNIV SCHOOL OF MEDICINE
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Patent Information

Authority / Receiving Office
US · United States
Patent Type
Applications(United States)
Current Assignee / Owner
THE JOHN HOPKINS UNIV SCHOOL OF MEDICINE
Publication Date
2005-07-14
Estimated Expiration
Not applicable · inactive patent

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Abstract

Genomic imprinting is a parent of origin-dependent gene silencing that involves marking of alleles in the germline and differential expression in somatic cells of the offspring. Imprinted genes and abnormal imprinting have been implicated in development, human disease, and embryonic stem cell transplantation. We have established a model system for genomic imprinting using pluripotent 8.5 d.p.c. mouse embryonic germ (EG) cell lines derived from an interspecific cross. We find that allele-specific imprinted gene expression has been lost in these cells. However, partial restoration of allele-specific silencing can occur for some imprinted genes after in vitro differentiation of EG cells into somatic cell lineages, indicating the presence of a gametic memory that is separable from allele-specific gene silencing. We have also generated a library containing most methylated CpG islands. A subset of these clones was analyzed and revealed a subdivision of methylated CpG islands into 4 distinct subtypes: CpG islands belonging to high copy number repeat families; unique CpG islands methylated in all tissues; unique methylated CpG islands that are unmethylated in the paternal germline; and unique CpG islands methylated in tumors. This approach identifies a methylome of methylated CpG islands throughout the genome.
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Description

[0001] This application claims the benefit of application Ser. Nos. 60 / 206,158 and 60 / 206,161 filed May 22, 2000.

[0002] This invention was made using funds from the U.S. government under a grant from the National Institutes of Health numbered CA65145. The U.S. government therefore retains certain rights in the invention.BACKGROUND OF THE INVENTION

[0003] Genomic imprinting is a parental origin-specific gene silencing that leads to differential expression of the two alleles of a gene in mammalian cells. Imprinting has attracted intense interest for several reasons: (i) Imprinting is by definition reversible and may be regulated over a large genomic domain (1). (ii) Imprinted genes and the imprinting mechanism itself are important in human birth defects and cancer (2). (iii) It has been suggested that imprinting cannot be reprogrammed without passage through the germline and thus constitutes a barrier to human embryonic stem cell transplantation (3).

[0004] Experimental studies of the...

Claims

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