Transgenic cattle produced by reverse-transcribed gene transfer in oocytes

AWS Chan, EJ Homan, LU Ballou… - Proceedings of the …, 1998 - National Acad Sciences
AWS Chan, EJ Homan, LU Ballou, JC Burns, RD Bremel
Proceedings of the National Academy of Sciences, 1998National Acad Sciences
A critical requirement for integration of retroviruses, other than HIV and possibly related
lentiviruses, is the breakdown of the nuclear envelope during mitosis. Nuclear envelope
breakdown occurs during mitotic M-phase, the envelope reforming immediately after cell
division, thereby permitting the translocation of the retroviral preintegration complex into the
nucleus and enabling integration to proceed. In the oocyte, during metaphase II (MII) of the
second meiosis, the nuclear envelope is also absent and the oocyte remains in MII arrest for …
A critical requirement for integration of retroviruses, other than HIV and possibly related lentiviruses, is the breakdown of the nuclear envelope during mitosis. Nuclear envelope breakdown occurs during mitotic M-phase, the envelope reforming immediately after cell division, thereby permitting the translocation of the retroviral preintegration complex into the nucleus and enabling integration to proceed. In the oocyte, during metaphase II (MII) of the second meiosis, the nuclear envelope is also absent and the oocyte remains in MII arrest for a much longer period of time compared with M-phase in a somatic cell. Pseudotyped replication-defective retroviral vector was injected into the perivitelline space of bovine oocytes during MII. We show that reverse-transcribed gene transfer can take place in an oocyte in MII arrest of meiosis, leading to production of offspring, the majority of which are transgenic. We discuss the implications of this mechanism both as a means of production of transgenic livestock and as a model for naturally occurring recursive transgenesis.
National Acad Sciences
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