Transgenic cattle produced by reverse-transcribed gene transfer in oocytes
Transgenic cattle produced by reverse-transcribed gene transfer in oocytes
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DOI：
10.1073/pnas.95.24.14028
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发表时间：
1998-11-24
影响因子：
11.1
通讯作者：
Bremel, RD
中科院分区：
文献类型：
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作者：
Chan, AWS;Homan, EJ;Bremel, RD
除了HIV和可能相关的慢病毒之外，逆转录病毒整合的关键要求是在有丝分裂期间核膜的破坏。核膜破裂发生在有丝分裂M期，细胞分裂后立即重新形成核膜，从而允许逆转录病毒整合前复合物易位到细胞核中并使整合得以进行。在卵母细胞中，在第二次减数分裂的中期II（MII）期间，核被膜也不存在，并且与体细胞中的M期相比，卵母细胞保持在MII停滞的时间长得多。将复制缺陷型逆转录病毒载体注射到牛卵母细胞的卵黄周隙中。我们发现，逆转录的基因转移可以发生在卵母细胞在MII逮捕减数分裂，导致生产的后代，其中大部分是转基因的。我们讨论了这一机制的影响，既作为生产转基因牲畜的一种手段，也作为自然发生的递归转基因的模型。
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.