A functional floxed allele of pkd1 that can be conditionally inactivated in vivo
A functional floxed allele of pkd1 that can be conditionally inactivated in vivo
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DOI：
10.1097/01.asn.0000144204.01352.86
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发表时间：
2004-12-01
影响因子：
13.6
通讯作者：
Germino, GG
中科院分区：
文献类型：
--
作者：
Piontek, KB;Huso, DL;Germino, GG
基因打靶已被用于创建具有 Pkd1 突变的多种 Trice 品系，这些突变具有许多共同特征。纯合 Pkd1 突变体如果存活到性交后 15.5 天，并且在胎儿或围产期死亡，则总是会出现胰腺和肾囊肿。时期。相比之下，携带 Pkd1 杂合突变的小鼠通常是正常的，并且几乎没有肾囊肿。这些特征限制了这些模型作为研究囊肿形成发病机制和各种治疗干预对疾病进展的影响的工具的实用性。该报告描述了具有 Pkd1 (Pkd1(cond)) 的 floxed 等位基因的新小鼠品系，其内含子 1 中插入有 FRT 侧翼的新霉素盒，内含子 1 和内含子 4 中插入有 lox P 位点。Pkd1(cond) 等位基因功能齐全，纯合子能够存活且健康。结果表明，通过分别与表达 cre 或 FLPe 重组酶的动物杂交，可以选择性诱导 lox P 和 FRT 位点重组，产生两个新的等位基因 Pkd1(del2-4) 和 Pkd1(cond-Deltaneo)。结果发现，Pkd1(del2-4) 等位基因的功能为真正的无效，而新霉素基因的存在或不存在则没有功能影响。研究还表明，Pkd1 的体细胞丢失会导致肾囊肿和肝囊肿。这种新的小鼠品系在 Pkd1 生物学研究中将具有无价的价值，并可作为一种强大的新工具，可用于研究常染色体显性多囊肾病的发病机制。
Gene targeting has been used to create a variety of lines of trice with Pkd1 mutations that share many common features. Homozygous Pkd1 mutants invariably develop pancreatic and renal cysts if they survive to day 15.5 post coitum and die in either the fetal or the perinatal. period. In contrast, mice with heterozygous mutations of Pkd1 are generally normal and have few if any renal cysts. These features have limited the utility of these models as tools to study the pathogenesis of cyst formation and the effect of various therapeutic interventions on disease progression. This report describes a new line of mice with a floxed allele of Pkd1 (Pkd1(cond)) that has an FRT-flanked neomycin cassette inserted into intron 1 and lox P sites inserted into intron 1 and intron 4. The Pkd1(cond) allele is fully functional, and homozygotes are viable and healthy. It is shown that the lox P and FRT sites can be selectively induced to recombine to produce two new alleles, Pkd1(del2-4) and Pkd1(cond-Deltaneo), by crossing to animals that express either the cre or FLPe recombinase, respectively. It is found that Pkd1(del2-4) allele functions as a true null, whereas presence or absence of the neomycin gene has no functional effects. It also is shown that somatic loss of Pkd1 results in renal and hepatic cysts. This new line of mice will be invaluable in the study of Pkd1 biology and serve as a powerful new tool that can be used to study the pathogenesis of autosomal dominant polycystic kidney disease.