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用于对经历了Cre介导的重组的细胞进行非侵入性生物发光成像的小鼠报告菌株。

Mouse reporter strain for noninvasive bioluminescent imaging of cells that have undergone Cre-mediated recombination.

作者信息

Safran Michal, Kim William Y, Kung Andrew L, Horner James W, DePinho Ron A, Kaelin William G

机构信息

Dana-Farber Cancer Institute and Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.

出版信息

Mol Imaging. 2003 Oct;2(4):297-302. doi: 10.1162/15353500200303154.

Abstract

Conditional alleles containing LoxP recombination sites, in conjunction with Cre recombinase delivered by a variety of means, allows for spatial and temporal control of gene expression in mouse models. Here we describe a mouse strain in which a luciferase (Luc) cDNA, preceded by a LoxP-stop-LoxP (L-S-L) cassette, was introduced into the ubiquitously expressed ROSA26 locus. Mouse embryo fibroblasts derived from this strain expressed luciferase after Cre-mediated recombination in vitro. ROSA26 L-S-L-Luc/+ mice expressed luciferase in a diffuse or liver-restricted pattern, as determined by noninvasive, bioluminescent imaging, when crossed to transgenic mice in which Cre was under the control of a zygotically expressed (EIIA-Cre), or a liver-restricted (albumin-Cre), promoter, respectively. Organ-specific luciferase expression was also seen after intraparenchymal administration of an adenovirus encoding Cre. The ROSA26 L-S-L-Luc/+ strain should be useful for characterizing Cre mouse strains and for following the fate of cells that have undergone Cre-mediated recombination in vivo.

摘要

含有LoxP重组位点的条件性等位基因,与通过多种方式递送的Cre重组酶相结合,能够在小鼠模型中实现基因表达的时空控制。在此,我们描述了一种小鼠品系,其中一个带有LoxP-stop-LoxP(L-S-L)盒的荧光素酶(Luc)cDNA被引入到广泛表达的ROSA26基因座中。源自该品系的小鼠胚胎成纤维细胞在体外经Cre介导的重组后表达荧光素酶。当与Cre分别受合子表达启动子(EIIA-Cre)或肝脏特异性启动子(白蛋白-Cre)控制的转基因小鼠杂交时,通过非侵入性生物发光成像确定,ROSA26 L-S-L-Luc/+小鼠以弥漫性或肝脏限制性模式表达荧光素酶。在实质内给予编码Cre的腺病毒后,也观察到了器官特异性荧光素酶表达。ROSA26 L-S-L-Luc/+品系应有助于鉴定Cre小鼠品系,并追踪体内经历Cre介导重组的细胞的命运。

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