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

页码：411 / 417

页数：7

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