Epigenetic markers of pluripotency
AssigneeZymo Research Corporation
InventorsJill PETRISKO, Lam NGUYEN, Manuel KRISPIN
Life of this patent
Estimating the term…
Abstract
Epigenetic methods for assessing pluripotency of a cell population, such as a stem cell culture are provided. For example, pluripotency can be assessed by determining DNA methylation status at the RAB25, NANOG, PTPN6, MGMT, GBP3 and/or LYST gene regions. Kits and reagents for testing cells are likewise provided.
Claims27
A method for assessing pluripotency in a cell population comprising:
- (a) obtaining a nucleic acid sample from the cell population;
- and
- (b) determining the DNA methylation status at 2 or more gene regions selected from the group consisting of RAB25, NANOG, PTPN6, MGMT, GBP3 and LYST,
- wherein an increased level of DNA methylation relative to a reference at MGMT, GBP3 or LYST;
- or a decreased level of DNA methylation relative to a reference at RAB25, NANOG or PTPN6 indicates that the cell population comprises pluripotent cells.
A kit comprising a sealed container comprising primers designed to amplify regions including potential DNA methylation sites in at least two gene regions selected from the group consisting of RAB25, NANOG, PTPN6, MGMT, GBP3 and LYST.
A method for assessing pluripotency in a cell population comprising:
- (a) obtaining a DNA sample from the cell population;
- (b) identifying two or more genomic amplification intervals, each interval comprising at least one CpG position within the recognition sequence of a methylation-sensitive endonuclease (MSRE), where the CpG position is subject to differential methylation during cell differentiation;
- (c) amplifying the two or more genomic amplification intervals in the presence and absence of the MSRE;
- and
- (d) quantifying the amount of amplification product to determine the proportion of DNA methylation in the two or more genomic amplification intervals, thereby assessing pluripotency in the cell population.
Dependencies are inferred from the claim text.