US 10,893,666 B2 · Production of fertile XY female animals by silencing of genes on the Y chromosome

Abstract

Methods and compositions are provided for generating F0 fertile XY female animals. The methods and compositions involve making XY pluripotent or totipotent animal cells, in vitro cell cultures, or embryos that are capable of producing a fertile female XY animal in an F0 generation. Such cells, embryos, and animals can be made by silencing a region of the Y chromosome. Optionally, the cells can also be cultured in feminizing medium such as a low-osmolality medium and/or can be modified to decrease the level and/or activity of an Sry protein. Methods and compositions are also provided for silencing a region of the Y chromosome in an XY pluripotent or totipotent animal cell, or in vitro cell cultures, embryos, or animals derived therefrom, by maintaining an XY pluripotent or totipotent animal cell in a feminizing medium. Methods and compositions are also provided for maintaining a population of XY pluripotent or totipotent animal cells in a feminizing medium and selecting cells or clones having increased capabilities for producing a fertile female XY animal in an F0 generation. Methods and compositions are also provided for screening for compounds with feminizing activity or for optimizing concentrations of components in feminizing media.

Claims26

Dependent claims

A method for screening a target compound for making a mouse XY embryonic stem (ES) cell capable of producing a fertile female XY mouse in an FO generation, said method comprising:

(a) culturing a first population and a second population of mouse XY ES cells in a medium comprising a base medium and supplements suitable for maintaining the pluripotency of the mouse XY ES cells but that does not alter the cells' capacity to give rise to fertile female progeny, wherein the first population and the second population originate from the same cell line or the same clone or have the same genotype, and wherein the first population is cultured in the presence of the target compound and the second population is cultured in the absence of the target compound;
(b) assaying one or more of the mouse XY ES cells in each of the first and second populations of mouse XY ES cells and detecting mRNA expression of Ddx3y, Uty, and Eif2s3y;
and
(c) selecting the target compound that decreases mRNA expression of Ddx3y, Uty, and Eif2s3y by at least 90% in the one or more mouse XY ES cells from the first population as compared to the one or more mouse XY ES cells from the second population, thereby selecting the target compound for making the mouse XY ES cell capable of producing the fertile female XY mouse in the FO generation.

1 more element

Dependencies are inferred from the claim text.

Production of fertile XY female animals by silencing of genes on the Y chromosome

Life of this patent

Production of fertile XY female animals by silencing of genes on the Y chromosome

Life of this patent

Abstract

Claims26