Spontaneous recombinase activity of Cre-ERT2 in vivo

被引：46
|
作者：
Kristianto, Jasmin [1 ,2 ]
Johnson, Michael G. [4 ]
Zastrow, Ryley K. [4 ]
Radcliff, Abigail B. [4 ]
Blank, Robert D. [1 ,2 ,3 ]
机构：
[1] Med Coll Wisconsin, Dept Med, Milwaukee, WI 53226 USA
[2] William S Middleton Mem Vet Adm Med Ctr, Madison, WI USA
[3] Milwaukee VA Med Ctr, Milwaukee, WI USA
[4] Univ Wisconsin, Dept Med, Sch Med & Publ Hlth, Madison, WI USA
来源：
TRANSGENIC RESEARCH | 2017年 / 26卷 / 03期
关键词：
Tamoxifen; Estrogen receptor; Recombination; Cre-activity; TETRACYCLINE-RESPONSIVE PROMOTERS; GENE-EXPRESSION; TIGHT CONTROL; MICE; MUTAGENESIS; VECTORS; CELLS;
D O I：
10.1007/s11248-017-0018-1
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Inducible Cre-ERT recombinase technology is widely used for gene targeting studies. The second generation of inducible Cre-ERT recombinase, hemizygous B6.129S-Tg(UBC-cre/ERT2)1Ejb/J (hereafter abbreviated as Cre-ERT2), a fusion of a mutated estrogen receptor and Cre recombinase, was engineered to be more efficient and specific than the original Cre-ERT. The putative mechanism of selective Cre-mediated recombination is Cre sequestration in the cytoplasm in the basal state with translocation to the nucleus only in the presence of tamoxifen. We utilized both a reporter mouse (B6.129 (Cg)-Gt(ROSA)26Sor (tm4(ACTB-tdTomato,-EGFP)Luo) /J) and endothelin converting enzyme-1 floxed transgenic mouse line to evaluate Cre-ERT2 activity. We observed spontaneous Cre activity in both settings. Unintended Cre activity is a confounding factor that has a potentially large impact on data interpretation. Thus, it is important to consider background Cre activity in experimental design.
引用
收藏
页码：411 / 417
页数：7
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