Qiu Min Ru, Jiang Lele, Matthaei Klaus I, Schoenwaelder Simone M, Kuffner Tamara, Mangin Pierre, Joseph Joanne E, Low Joyce, Connor David, Valenzuela Stella M, Curmi Paul M G, Brown Louise J, Mahaut-Smith Martyn, Jackson Shaun P, Breit Samuel N
St. Vincent's Centre for Applied Medical Research, St. Vincent's Hospital and University of New South Wales, Sydney, New South Wales, Australia.
Genesis. 2010 Feb;48(2):127-36. doi: 10.1002/dvg.20590.
CLIC1 belongs to a family of highly conserved and widely expressed intracellular chloride ion channel proteins existing in both soluble and membrane integrated forms. To study the physiological and biological role of CLIC1 in vivo, we undertook conditional gene targeting to engineer Clic1 gene knock-out mice. This represents creation of the first gene knock-out of a vertebrate CLIC protein family member. We first generated a Clic1 Knock-in (Clic1(FN)) allele, followed by Clic1 knock-out (Clic1(-/-)) mice by crossing Clic1(FN) allele with TNAP-cre mice, resulting in germline gene deletion through Cre-mediated recombination. Mice heterozygous or homozygous for these alleles are viable and fertile and appear normal. However, Clic1(-) (/-) mice show a mild platelet dysfunction characterized by prolonged bleeding times and decreased platelet activation in response to adenosine diphosphate stimulation linked to P2Y(12) receptor signaling.
CLIC1属于一个高度保守且广泛表达的细胞内氯离子通道蛋白家族，以可溶性和膜整合形式存在。为了研究CLIC1在体内的生理和生物学作用，我们进行了条件性基因靶向操作，构建了Clic1基因敲除小鼠。这是首次对脊椎动物CLIC蛋白家族成员进行基因敲除。我们首先构建了一个Clic1敲入（Clic1(FN)）等位基因，然后通过将Clic1(FN)等位基因与TNAP-cre小鼠杂交，获得了Clic1敲除（Clic1(-/-)）小鼠，通过Cre介导的重组实现了种系基因缺失。这些等位基因的杂合或纯合小鼠均可存活且可育，外观正常。然而，Clic1(-/-)小鼠表现出轻度血小板功能障碍，其特征是出血时间延长，并且在与P2Y(12)受体信号传导相关的二磷酸腺苷刺激下血小板活化降低。
