Methods for mitochondrial DNA replacement in oocytes

摘要

Methods are provided for producing a primate oocyte in vitro. The methods include removing nuclear DNA from a recipient primate oocyte from a first primate in a manner that does not lower levels of maturation promoting factor (MPF) to form an enucleated recipient primate oocyte. The recipient primate oocyte is enucleated using a non-UV-based spindle imaging system. Nuclear genetic material or DNA including chromosomes from a donor primate oocyte arrested at metaphase II from a second primate is isolated in the form of the karyoplast and introduced into the enucleated recipient primate oocyte. Introduction of the chromosomes is performed using a fusogenic agent or electroporation to produce a hybrid oocyte.

主权项

1. A method for producing an oocyte in vitro comprising the steps of:
(a) enucleating a recipient primate oocyte arrested at metaphase II from a first primate in a manner that does not lower levels of maturation promoting factor (MPF) to form an enucleated recipient primate oocyte, wherein the recipient primate oocyte is enucleated using a non-UV-based spindle imaging system; and (b) isolating nuclear genetic material comprising chromosomes from a donor primate oocyte arrested at metaphase II from a second primate and introducing into the enucleated recipient primate oocyte, wherein introduction of the chromosomes is performed using a fusogenic agent, wherein the fusogenic agent is Sendai virus, and wherein the first primate and the second primate are from the same primate species, thereby producing a hybrid oocyte arrested at metaphase II, wherein the hybrid oocyte can be can be fertilized.