FACTORS AFFECTING THE EFFICIENCY OF INTRODUCING FOREIGN DNA INTO MICE BY MICROINJECTING EGGS
FACTORS AFFECTING THE EFFICIENCY OF INTRODUCING FOREIGN DNA INTO MICE BY MICROINJECTING EGGS
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DOI：
10.1073/pnas.82.13.4438
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
PALMITER, RD
中科院分区：
文献类型：
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作者：
BRINSTER, RL;CHEN, HY;PALMITER, RD
将外源DNA显微注射到哺乳动物受精卵中，是将基因引入生殖系的一种方便的方法。描述了影响外源DNA成功整合到小鼠染色体中的一些更重要的参数。DNA浓度、大小和形状(超螺旋和线形，末端不同)以及注射部位(男性原核、女性原核或细胞质)和缓冲液成分的影响都被考虑在内。整合的最佳条件需要将数百个线性分子注入受精卵的雄性原核。在这些条件下，.apprx。发育成熟的小鼠中有25%继承了一份或多份显微注射的DNA。总体效率还取决于小鼠品系的选择；例如，产生表达外源生长激素基因的转基因小鼠是.apprx。使用C57/BL6的速度提高了8倍。SJL杂交鼠优于近交系C57/BL6小鼠。
Microinjection of foreign DNA into fertilized mammalian eggs is a convenient means of introducing genes into the germ line. Some of the more important parameters that influence successful integration of foreign DNA into mouse chromosomes are described. The effects of DNA concentration, size, and form (supercoiled vs. linear with a variety of different ends) are considered as well as the site of injection (male pronucleus, female pronucleus or cytoplasm) and buffer composition. The optimal conditions for integration entail injection of a few hundred linear molecules into the male pronucleus of fertilized 1 cell eggs. Under these conditions .apprx. 25% of the mice that develop inherit 1 or more copies of the microinjected DNA. The overall efficiency also depends on the choice of mouse strains; for example, generating transgenic mice that express foreign growth hormone genes is .apprx. 8 times easier with C57/BL6 .times. SJL hybrid mice than with inbred C57/BL6 mice.