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US 7,411,111 B2Granted
Method for cloning of the rat by nuclear transfer
AssigneeInstitut National de la Recherche Agronomique
InventorsJean Cozzi, Qi Zhou, Jean-Paul Renard
Life of this patent
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Abstract
The invention relates to a method for cloning in the rat by nuclear transfer. The invention further relates to the rats obtained thus, in the foetal or adult state, as well as use thereof for the production of molecules of interest or as study models.
Claims38
A method for producing of viable reconstituted rat embryo by nuclear transfer comprising the following steps:
- a) removing a recipient rat ovocyte and maintaining the recipient _ovocyte in vitro in a maintenance medium, wherein the recipient rat ovocyte is maintained at the metaphase stage of the second meiotic division in a maintenance medium including a reversible proteasome inhibitor sufficient to prevent activation of the ovocyte;
- b) transferring the nucleus of a rat donor cell into the recipient rat ovocyte of step a);
- c) removing the recipient rat ovocyte nucleus before or after the introduction of said donor cell nucleus to obtain a reconstituted rat ovocyte;
- and
- d) activating the reconstituted rat ovocyte by placing the reconstituted ovocyte in an activation medium containing an inducer of activation to obtain a reconstituted rat embryo.
2 more elements
A method for producing of cloned rat foetus by nuclear transfer comprising the following steps:
- a) removing a recipient rat ovocyte and maintaining the recipient_ovocyte in vitro in a maintenance medium, wherein the recipient rat ovocyte is maintained at the metaphase stage of the second meiotic division in a maintenance medium including a reversible proteasome inhibitor sufficient to prevent activation of the ovocyte;
- b) transferring the nucleus of a rat donor cell into the recipient rat ovocyte of step a);
- c) removing the recipient rat ovocyte nucleus before or after the introduction of said donor cell nucleus to obtain a reconstituted rat ovocyte;
- d) activating the reconstituted rat ovocyte by placing the reconstituted ovocyte in an activation medium containing an inducer of activation to obtain an activated reconstituted rat embryo;
- and
- e) transferring the activated reconstituted embryo into the oviduct or uterus of a recipient female rat wherein said transferred embryo is allowed to become a fetus.
3 more elements
A method for producing a cloned rat by nuclear transfer comprising the following steps:
- a) removing a recipient rat ovocyte and maintaining the recipient_ovocyte in vitro in a maintenance medium, wherein the recipient rat ovocyte is maintained at the metaphase stage of the second meiotic division in a maintenance medium including a reversible proteasome inhibitor sufficient to prevent activation of the ovocyte;
- b) transferring the nucleus of a rat donor cell into the recipient rat ovocyte of step a);
- c) removing the recipient rat ovocyte nucleus before or after the introduction of said donor cell nucleus to obtain a reconstituted rat ovocyte;
- d) activating the reconstituted rat ovocyte by placing the reconstituted ovocyte in an activation medium containing an inducer of activation to obtain an activated reconstituted rat embryo;
- e) transferring the activated reconstituted embryo into the oviduct or uterus of a recipient female rat wherein said transferred embryo is allowed to become implanted and to develop into a cloned rat in the uterus of said recipient female;
- and
- f) recovering a neonate at birth.
4 more elements
Dependencies are inferred from the claim text.