O'Neal Kelly R, Agah Ramtin
The Program in Human Molecular Biology and Genetics, University of Utah, Salt Lake City, USA.
Methods Mol Biol. 2007;366:309-20. doi: 10.1007/978-1-59745-030-0_17.
Tissue specific gene knockouts using Cre recombinase can have broad applicability in murine disease models of cardiovascular disease. The Cre system has been shown to have broad experimental versatility for both temporal and spatial control of gene deletion. By and large this is achieved by first generating mice with an inducible tissue specific promoter for expression of Cre. These mice can then be crossed with a second line of mice where the gene of interest in 'knocked in' flanked by Cre recognition sequences Lox-P sites. The double transgenic lines are then induced, through administration of an exogenous agent, to allow tissue specific, i.e. cardiac, knockout of the gene of interest at the desired time. An experimental protocol delineating this technique is described in the chapter.
使用Cre重组酶进行组织特异性基因敲除在心血管疾病的小鼠疾病模型中具有广泛的适用性。Cre系统已被证明在基因缺失的时间和空间控制方面具有广泛的实验通用性。大体上,这是通过首先产生具有可诱导的组织特异性启动子以表达Cre的小鼠来实现的。然后可以将这些小鼠与第二系小鼠杂交,在第二系小鼠中,感兴趣的基因被“敲入”,两侧为Cre识别序列Lox-P位点。然后通过给予外源性试剂诱导双转基因系,以在所需时间实现感兴趣基因的组织特异性(即心脏)敲除。本章描述了描述该技术的实验方案。