Bader Michael
Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany.
Methods Mol Med. 2005;108:17-32. doi: 10.1385/1-59259-850-1:017.
Gene-targeting technology allows the planned alteration of any gene in the mouse genome and has been very successfully employed to study the function of numerous gene products in a complete animal. The method includes the design of a suitable targeting construct, its transfection into pluripotential embryonic stem cells, selection for cells in which one allele of the endogenous gene has been exchanged for the construct by homologous recombination, and the transfer of these cells into host blastocysts. The blastocysts are then transferred into the uterus of a pseudopregnant foster mother giving birth to chimeras, which are bred to yield heterozygous and finally homozygous mutant mice for the targeted gene.
基因靶向技术能够对小鼠基因组中的任何基因进行有计划的改造，并且已非常成功地用于在完整动物体内研究众多基因产物的功能。该方法包括设计合适的靶向构建体，将其转染到多能胚胎干细胞中，筛选出通过同源重组使内源基因的一个等位基因被构建体替换的细胞，然后将这些细胞转移到宿主囊胚中。接着将囊胚转移到假孕代孕母鼠的子宫内，产出嵌合体，再将嵌合体进行繁殖，以获得针对靶向基因的杂合子，最终获得纯合子突变小鼠。
