Methods of analysis of allelic imbalance
View Patent ↗Methods are provided for identification of genes that are imprinted. In another embodiment methods are provided for identification and analysis of genes whose expression shows allelic imbalance. The expression products transcribed from genes that are present in the genome as two or more alleles may be distinguished by hybridization to an array designed to interrogate individual alleles. Genes whose transcription products are present in amounts that vary from expected are candidates for allelic imbalance, imprinting and imprinting errors.
1. A method for determining if a patient has an increased risk of developing a disease due to abnormal imprinting, comprising the steps of:
(a) providing a nucleic acid array comprising probes designed to interrogate the genotype of a plurality of single nucleotide polymorphisms in genes that are known to be imprinted and to show monoallelic expression in healthy individuals;
(b) providing a genomic DNA sample from the patient and labeling said genomic DNA sample with a first detectable label;
(c) isolating an RNA sample from the patient and labeling said RNA sample with a second detectable label, wherein said first and second detectable labels are separately detectable;
(d) hybridizing the genomic DNA sample labeled with said first detectable label and the RNA sample labeled with said second detectable label to the nucleic acid array;
(e) generating a hybridization pattern resulting from said hybridization and analyzing the hybridization pattern to determine the genotype of a plurality of single nucleotide polymorphisms in the genomic DNA and to determine the genotype of the same single nucleotide polymorphisms in the RNA, to identify one or more single nucleotide polymorphisms in the plurality of single nucleotide polymorphisms that are heterozygous in the genomic DNA and heterozygous in the RNA sample from the patient and thereby identifying a gene that is showing biallelic expression in the patient; and
(f) identifying the patient as having an increased risk of developing a disease if one or more single nucleotide polymorphisms that show monoallelic expression in healthy individuals is identified as showing biallelic expression in the patient.
2. The method of claim 1 wherein the disease is cancer.
3. The method of claim 1 wherein the disease is a neurological disorder.
4. The method of claim 1 wherein the array includes probes to interrogate the genotype of genes that encode a lymphoid-specific factor, an olfactory receptor, a T cell receptor, and a subunit of an immunoglobulin.
5. The method of claim 1 wherein said plurality of single nucleotide polymorphisms comprises at least 1,000 different polymorphic positions.
6. The method of claim 1 wherein the array includes probes to genotype single nucleotide polymorphisms in at least 100 genes that are known to be imprinted.