Dnmt3L-related cloning and reproductive methods
OwnerTimothy H. Bestor, Deborah Bourc'hisinventors
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Abstract
This invention provides methods for reducing the amount of Dnmt3L in a cell, obtaining a cell having a reduced amount of Dnmt3L, and cells, tissues, organs and non-human transgenic mammals having a reduced amount of Dnmt3L. This invention further provides related methods for producing a mammalian zygote in vitro which, upon successful subsequent development in utero, gives rise to a mammal having a reduced susceptibility to an abnormality associated with epigenetic instability. This invention further provides methods for determining the amount of Dnmt3L in a cell, and methods for determining whether agents bind to Dnmt3L or decrease the amount of Dnmt3L in a cell. Finally, this invention provides related nucleic acids and compositions.
Claims48
A method for reducing the amount of Dnmt3L in a cell comprising contacting the cell under suitable conditions with an agent that specifically inhibits the expression of Dnmt3L in the cell, thereby reducing the amount of Dnmt3L in the cell.
A method for obtaining a cell having a reduced amount of Dnmt3L comprising isolating a cell from a transgqnic non-human mammal whose cells contain a reduced amount of Dnmt3L.
An isolated cell having a reduced amount of Dnmt3L.
An isolated population of cells having a reduced amount of Dnmt3L.
A tissue comprising cells having a reduced amount of Dnmt3L.
An organ comprising cells having a reduced amount of Dnmt3L.
A non-human transgenic mammal comprising cells having a reduced amount of Dnmt3L.
A method for producing a mammalian zygote in vitro which, upon successful subsequent development in utero, gives rise to a mammal having a reduced susceptibility to an abnormality associated with epigenetic instability, comprising contacting a mammalian sperm cell and a mammalian ovum in vitro under conditions permitting fertilization of the ovum by the sperm, wherein the sperm cell, the ovum or both are contacted with an agent that reduces the amount of Dnmt3L therein, thereby producing the mammalian zygote.
A method for producing a Dnmt3L− non-human mammalian cell, which method comprises introducing a non-human mammalian Dnmt3L nucleus into a non-human mammalian enucleated oocyte or zygote under suitable conditions, thereby producing a Dnmt3L− cell.
A method for producing a non-human mammalian zygote which, upon successful subsequent development in utero, gives rise to a non-human mammal having a reduced susceptibility to an abnormality associated with epigenetic instability, comprising introducing a non-human mammalian Dnmt3L− nucleus into a non-human mammalian enucleated oocyte or zygote under suitable conditions, thereby producing the zygote.
A method for determining the amount of Dnmt3L in a cell comprising (a) contacting the cell's contents with an agent that specifically forms a complex with Dnmt3L, (b) determining the amount of complex formed in step (a), and (c) comparing the amount of complex so determined with a known standard, thereby determining the amount of Dnmt3L in the cell.
A method for determining the amount of Dnmt3L in a cell comprising (a) determining the amount of Dnmt3L-encoding mRNA in the cell and (b) comparing the amount so determined with a known standard permitting the quantitation of Dnmt3L based on a known amount of Dnmt3L-encoding mRNA, thereby determining the amount of Dnmt3L in the cell.
A method for determining whether an agent binds to Dnmt3L comprising contacting the agent with Dnmt3L under suitable conditions, and determining whether the agent forms a complex with the Dnmt3L, the formation of a complex indicating that the agent binds to Dnmt3L.
A method for determining whether an agent decreases the amount of Dnmt3L in a cell, which method comprises the steps of:
- (a) contacting the cell with the agent under suitable conditions;
- (b) determining the amount of Dnmt3L in the cell after a suitable period of time;
- and
- (c) comparing the amount of Dnmt3L determined in step (b) with the amount of Dnmt3L in a cell in the absence of the agent, a lower amount of Dnmt3L in the cell contacted with the agent indicating that the agent decreases the amount of Dnmt3L in the cell.
A method for determining whether an agent decreases the expression of Dnmt3L in a cell, which method comprises the steps of:
- (a) contacting the cell with the agent under suitable conditions;
- (b) determining the amount of Dnmt3L expression in the cell after a suitable period of time;
- and
- (c) comparing the amount of Dnmt3L expression determined in step (b) with the amount of Dnmt3L expression in a cell in the absence of the agent, a lower amount of Dnmt3L expression in the cell contacted with the agent indicating that the agent decreases the amount of Dnmt3L expression in the cell.
An isolated nucleic acid that specifically hybridizes to Dnmt3L-encoding mRNA.
Dependencies are inferred from the claim text.