US5496720A
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Parthenogenic oocyte activation
Assignee: PROTEIN GENETICS Inc
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Filing: 1993-02-10
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Granted: 1996-03-05
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Expires: 2013-02-10
Abstract
Abstract A process of parthenogenic activation of mammalian oocytes which includes increasing intercellular levels of divalent cations in the oocyte; and reducing phosphorylation of cellular proteins in the oocyte. One method of accomplishing this is by introducing Ca 2+ free cation, such as ionomycin, to the oocyte and then preventing phosphorylation of the cellular proteins within the oocyte by adding a serine-threonine kinase inhibitor, such as 6-dimethylaminopurine (DMAP).
Inventors
- 👤 Joan L. Susko-Parrish
- 👤 David L. Northey
- 👤 M. Lorraine Leibfried-Rutledge
- 👤 Steven L. Stice
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 09:52
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/873; C12N5/00; C12N5/06; C12N5/0602; C12N5/0608; C12N5/0609; C12N2500/00; C12N2500/05; C12N2500/10; C12N2500/12; C12N2500/14; C12N2501/00; C12N2501/70; C12N2501/999
Claims Analysis
Total Claims: 28
Claim 1
Independent Claim
38 words
A process for the in vitro parthenogenic activation of a bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cations in the oocyte; and b. reducing phosphorylation of cellular proteins in the oocyte.
Claim 2
Dependent Claim
38 words
A process for the in vitro parthenogenic activation of a bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cations in the oocyte; and b. reducing phosphorylation of cellular proteins in the oocyte.
Claim 3
Dependent Claim
23 words
The process of claim 1 wherein the intracellular levels of divalent cation are increased by introducing a divalent cation into the oocyte cytoplasm.
Claim 4
Dependent Claim
21 words
The process of claim 2 wherein the divalent cation is selected from the group consisting of magnesium, strontium, barium and calcium.
Claim 5
Dependent Claim
14 words
The process of claim 2 wherein the divalent cation is a free calcium ion.
Claim 6
Dependent Claim
14 words
The process of claim 2 wherein the divalent cation is introduced by an ionophore.
Claim 7
Dependent Claim
18 words
The process of claim 5 wherein the ionophore is selected from the group consisting of ionomycin and A23187.
Claim 8
Dependent Claim
17 words
The process of claim 1 wherein the intracellular levels of divalent cation are increased by electric shock.
Claim 9
Dependent Claim
18 words
The process of claim 1 wherein the intracellular levels of divalent cation are increased by treatment with ethanol.
Claim 10
Dependent Claim
19 words
The process of claim 1 wherein the intracellular levels of divalent cation are increased by treatment with caged chelators.
Claim 11
Dependent Claim
15 words
The process of claim 1 wherein phosphorylation of cellular proteins is reduced by inhibiting phosphorylation.
Claim 12
Dependent Claim
26 words
The process of claim 10 wherein the process of inhibiting phosphorylation comprises adding an effective phosphorylation inhibiting amount of a serine-threonine kinase inhibitor to the oocyte.
Claim 13
Dependent Claim
22 words
The process of claim 11 wherein the serine-threonine kinase inhibitor is selected from the group consisting of 6-dimethylaminopurine, staurosporine, 2-aminopurine, and sphingosine.
Claim 14
Dependent Claim
12 words
The process of claim 11 wherein the serine-threonine kinase inhibitor is 6-dimethylaminopurine.
Claim 15
Dependent Claim
18 words
The process of claim 1 wherein phosphorylation of cellular proteins is reduced by inducing dephosphorylation in the oocyte.
Claim 16
Dependent Claim
13 words
The process of claim 1 wherein the oocyte is a 10-52 hour oocyte.
Claim 17
Dependent Claim
13 words
The process of claim 1 wherein the oocyte is a 16-30 hour oocyte.
Claim 18
Dependent Claim
14 words
The process of claim 1 wherein the oocyte is is approximately 24 hours old.
Claim 19
Dependent Claim
72 words
A process for the in vitro parthenogenic activation of a 10-52 hour bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cation in the oocyte by introducing a divalent cation into the oocyte cytoplasm; and b. reducing phosphorylation of cellular proteins in the oocyte wherein phosphorylation of cellular proteins is reduced by adding a serine-threonine kinase inhibitor to the oocyte in an amount effective to inhibit phosphorylation.
Claim 20
Dependent Claim
72 words
A process for the in vitro parthenogenic activation of a 10-52 hour bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cation in the oocyte by introducing a divalent cation into the oocyte cytoplasm; and b. reducing phosphorylation of cellular proteins in the oocyte wherein phosphorylation of cellular proteins is reduced by adding a serine-threonine kinase inhibitor to the oocyte in an amount effective to inhibit phosphorylation.
Claim 21
Dependent Claim
21 words
The process of claim 18 wherein the divalent cation is selected from the group consisting of magnesium, strontium, barium and calcium.
Claim 22
Dependent Claim
14 words
The process of claim 18 wherein the divalent cation is a free calcium ion.
Claim 23
Dependent Claim
12 words
The process of claim 18 wherein the serine-threonine kinase inhibitor is 6-dimethylaminopurine.
Claim 24
Dependent Claim
13 words
The process of claim 18 wherein the oocyte is a 16-30 hour oocyte.
Claim 25
Dependent Claim
14 words
The process of claim 18, wherein the oocyte is is approximately 24 hours old.
Claim 26
Dependent Claim
49 words
A process for the in vitro parthenogenic activation of a bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cation in the oocyte; and b. reducing phosphorylation of cellular proteins in the oocyte by introducing a dephosphorylizing amount of a phosphatase to the oocyte.
Claim 27
Dependent Claim
49 words
A process for the in vitro parthenogenic activation of a bovine oocyte comprising the following steps in sequence: a. increasing intracellular levels of divalent cation in the oocyte; and b. reducing phosphorylation of cellular proteins in the oocyte by introducing a dephosphorylizing amount of a phosphatase to the oocyte.
Claim 28
Dependent Claim
20 words
The process of claim 24 wherein the phosphatase is selected from the group consisting of Phosphatase 2A and Phosphatase 2B.
References Cited (Backward Citations)
US4994384A
Cited
US5096822A
Cited
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