Selective oxidation of 5-methylcytosine by tet-family proteins
AssigneesChildren's Medical Center Corporation, The United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsAnjana RAO, Mamta Tahiliani, Kian Peng Koh, Suneet Agarwal, Aravind Iyer
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Abstract
The present invention provides for novel methods for regulating and detecting the cytosine methylation status of DNA. The invention is based upon identification of a novel and surprising catalytic activity for the family of TET proteins, namely TET1, TET2, TET3, and CXXC4. The novel activity is related to the enzymes being capable of converting the cytosine nucleotide 5-methylcytosine into 5-hydroxymethylcytosine by hydroxylation.
Claims20
A method comprising contacting a nucleic acid molecule with an isolated enzyme in an amount effective to convert a methylated base of said nucleic acid molecule to an oxidized derivative thereof.
A kit comprising the following separate components:
- (a) at least one isolated enzyme capable of converting a methylated base of a nucleic acid molecule to an oxidized derivative thereof;
- and
- (b) packaging materials and instructions therein to use said kit to convert a methylated base of said nucleic acid molecule to said oxidized derivative thereof.
A method for determining familial predisposition to a myelodysplastic syndrome, a myeloproliferative disorder, acute myelogenous leukemia, systemic mastocytosis, or chronic myelomonocytic leukemia in an individual in need thereof, the method comprising (i) determining a level of 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof in CD34+ cells from an individual in need thereof, (ii) determining a level of 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof, in CD34+ cells from a family member of the individual, wherein the family member is affected with a myelodysplastic syndrome, a myeloproliferative disorder, acute myelogenous leukemia, systemic mastocytosis, or chronic myelomonocytic leukemia, and (iii) comparing the level of 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof in the CD34+ cells from the individual in need thereof with the level of 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof, in the CD34+ cells from the affected family member, wherein a decrease in the level of 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof, in the individual relative to the 5-methylcytsosine, 5-hydroxymethylcytsosine, or a combination thereof level in the affected family member is indicative of the individual being predisposed to a myelodysplastic syndrome, a myeloproliferative disorder, acute myelogenous leukemia, systemic mastocytosis, or chronic myelomonocytic leukemia.
Dependencies are inferred from the claim text.