Cre-ERT2在体内的自发重组酶活性。
Spontaneous recombinase activity of Cre-ERT2 in vivo.
作者信息
Kristianto Jasmin, Johnson Michael G, Zastrow Ryley K, Radcliff Abigail B, Blank Robert D
机构信息
Department of Medicine, Medical College of Wisconsin, Milwaukee, WI, USA.
William S. Middleton Memorial Veterans Hospital, Madison, WI, USA.
出版信息
Transgenic Res. 2017 Jun;26(3):411-417. doi: 10.1007/s11248-017-0018-1. Epub 2017 Apr 13.
Abstract
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 /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.
摘要
诱导型Cre-ERT重组酶技术广泛应用于基因靶向研究。第二代诱导型Cre-ERT重组酶，半合子B6.129S-Tg(UBC-cre/ERT2)1Ejb/J（以下简称为Cre-ERT2），是一种突变雌激素受体与Cre重组酶的融合体，经设计后比原始的Cre-ERT更高效、更具特异性。选择性Cre介导重组的推测机制是，在基础状态下Cre隔离于细胞质中，仅在存在他莫昔芬时才易位至细胞核。我们利用报告基因小鼠（B6.129 (Cg)-Gt(ROSA)26Sor /J）和内皮素转换酶-1基因敲除转基因小鼠品系来评估Cre-ERT2的活性。我们在两种情况下均观察到了自发的Cre活性。意外的Cre活性是一个混杂因素，对数据解释可能有很大影响。因此，在实验设计中考虑背景Cre活性很重要。
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