Compositions and methods for generating physiological X chromosome inactivation
AssigneeChildren's Medical Center Corporation
InventorsYi Zhang, Azusa Inoue
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Abstract
The present invention features compositions and methods for recapitulating physiological X-chromosome inactivation (XCI) in a cell, including a cell of any embryo generated by Somatic Cell Nuclear Transfer (SCNT). In one aspect, the invention features a method for generating physiological X chromosome inactivation in an embryo generated by SCNT, the method comprising injecting the embryo generated via SCNT with an H3K27me3-specific demethylase polypeptide or a polynucleotide encoding said demethylase. Disclosed herein are methods, compositions, and kits comprising an agent which increases the expression of genes encoding an H3K27me3-specific demethylase, or increases the activity of human H3K27me3-specific demethylase.
Claims13
A method for depleting and detecting maternal H3K27me3 at an imprinted X-inactivation-specific transcript (Xist) locus of a maternal X chromosome and inducing maternal Xist derepression and maternal X chromosome inactivation (XCI) in a mammalian embryo generated by Somatic Cell Nuclear Transfer (SCNT), the method comprising:
- injecting the embryo generated via SCNT with an H3K27me3-specific demethylase Kdm6b polypeptide comprising the amino acid sequence as set forth in SEQ ID NO:
- 2 and having H3K27me3-specific demethylase activity or a polynucleotide encoding said Kdm6b polypeptide;
- and
- detecting in the Kdm6b-injected embryo a down-regulated expression level of maternal X-linked genes compared to the levels of autosomal genes and genes that escape X-chromosome inactivation;
- wherein
- (i) the Kdm6b-injected embryo exhibits loss of the maternal H3K27me3 domain at the Xist locus;
- and
- (ii) maternal allele expression bias of X-linked genes is greater than 35-60% in the injected embryo compared with an embryo not injected with the Kdm6b polypeptide or with a polynucleotide encoding the Kdm6b polypeptide.
A method for depleting and detecting maternal H3K27me3 at an imprinted X-inactivation-specific transcript (Xist) locus of a maternal X chromosome and inducing maternal Xist derepression and maternal X chromosome inactivation (XCI) in a mammalian embryo generated by Somatic Cell Nuclear Transfer (SCNT), the method comprising:
- injecting the embryo generated via SCNT with an H3K27me3-specific demethylase Kdm6b polypeptide comprising an amino acid sequence with at least 98% identity to SEQ ID NO:
- 2 and having H3K27me3-specific demethylase activity or a polynucleotide encoding said Kdm6b polypeptide;
- and
- detecting in the Kdm6b-injected embryo a down-regulated expression level of maternal X-linked genes compared to the levels of autosomal genes and genes that escape X-chromosome inactivation;
- wherein
- (i) the Kdm6b-injected embryo exhibits loss of the maternal H3K27me3 domain at the Xist locus;
- and
- (ii) maternal allele expression bias of X-linked genes is greater than 35-60% in the injected embryo compared with an embryo not injected with the Kdm6b polypeptide or with a polynucleotide encoding the Kdm6b polypeptide.
Dependencies are inferred from the claim text.