FATE OF MICROINJECTED GENES IN PREIMPLANTATION MOUSE EMBRYOS
FATE OF MICROINJECTED GENES IN PREIMPLANTATION MOUSE EMBRYOS
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DOI：
10.1002/mrd.1080330410
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发表时间：
1992-12-01
影响因子：
2.5
通讯作者：
WALL, RJ
中科院分区：
文献类型：
--
作者：
BURDON, TG;WALL, RJ
利用聚合酶链反应（PCR）技术，对显微注射到小鼠胚胎中的基因从单细胞期到囊胚期的状态进行了跟踪。显微注射的DNA中检测到所有的一个，两个，四个细胞注射胚胎和44%的桑椹胚和26%的囊胚。头-尾连接的显微注射的基因，稳定整合的转基因阵列的一个共同特征，在所有胚胎中检测到显微注射的基因注射后，发生在注射的基因的末端的结构无关。显微注射的DNA的甲基化依赖性限制性内切酶Dpn我的敏感性在所有胚胎中丢失的两个细胞阶段（24小时），表明DNA甲基化的变化，独立的转基因整合。在致密化之前分裂球的解离揭示了胚胎内显微注射的DNA的镶嵌分布，并支持注射的基因形成有限数量的阵列的概念，这些阵列独立地分离，直到它们整合到基因组中或被降解。
The state of genes microinjected into mouse embryos was followed from the one-cell to the blastocyst stage using the polymerase chain reaction (PCR). Microinjected DNA was detected in all one-, two-, and four-cell injected embryos and in 44% of morula and 26% of blastocysts. Head-to-tail ligation of microinjected genes, a common feature of stably integrated transgene arrays, was detected in all embryos after injection of microinjected genes and occurred irrespective of the structure at the ends of the injected genes. Sensitivity of microinjected DNA to a methylation-dependent restriction endonuclease Dpn I was lost in all embryos by the two-cell stage (24 hr), indicating a change in DNA methylation, independent of transgene integration. Dissociation of blastomeres prior to compaction revealed a mosaic distribution of the microinjected DNA within the embryo and supports the notion that injected genes form a limited number of arrays, which segregate independently until they integrate into the genome or are degraded.