Engineering mouse chromosomes with Cre-loxP:: Range, efficiency, and somatic applications
Engineering mouse chromosomes with Cre-loxP:: Range, efficiency, and somatic applications
复制标题
DOI：
10.1128/mcb.20.2.648-655.2000
复制
发表时间：
2000-01-01
影响因子：
5.3
通讯作者：
Bradley, A
中科院分区：
文献类型：
--
作者：
Zheng, BH;Sage, M;Bradley, A
染色体重排是遗传学研究的重要资源。最近，已经建立了一种基于Cre-loxP的方法，将定义的染色体重排（缺失、重复和倒位）引入小鼠基因组（染色体工程）。为了系统地探索该技术的局限性，我们在小鼠11号染色体上评估了该策略。尽管当两个端点在同一染色体上时，Cre-loxP介导的重组效率随着遗传距离的增加而降低，但即使当遗传距离最大化时，效率也不受限制，在小鼠胚胎干细胞（ES）中，已经构建了包含多达四分之三的11号染色体的重排。虽然较大的缺失可能导致ES细胞死亡，但较小的缺失可以在ES细胞中和体内以组织或细胞类型特异性方式非常有效地产生。我们的结论是，任何染色体重排可以在ES细胞与Cre-loxP策略，只要它不影响细胞活力，体内染色体工程可以潜在地用于实现杂合性的体细胞损失，在创建人类癌症的小鼠模型。
Chromosomal rearrangements are important resources for genetic studies. Recently, a Cre-loxP-based method to introduce defined chromosomal rearrangements (deletions, duplications, and inversions) into the mouse genome (chromosome engineering) has been established. To explore the limits of this technology systematically, we have evaluated this strategy on mouse chromosome 11, Although the efficiency of Cre-loxP-mediated recombination decreases with increasing genetic distance when the two endpoints are on the same chromosome, the efficiency is not limiting even when the genetic distance is maximized, Rearrangements encompassing up to three quarters of chromosome 11 have been constructed in mouse embryonic stem (ES) cells. While larger deletions may Lead to ES cell lethality, smaller deletions can be produced very efficiently both in ES cells and in vivo in a tissue- or cell-type-specific manner. We conclude that any chromosomal rearrangement can be made in ES cells with the Cre-loxP strategy provided that it does not affect cell viability, In vivo chromosome engineering can be potentially used to achieve somatic losses of heterozygosity in creating mouse models of human cancers.