Attribution of vascular phenotypes of the murine Egfl7 locus to the microRNA miR-126
Attribution of vascular phenotypes of the murine Egfl7 locus to the microRNA miR-126
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DOI：
10.1242/dev.029736
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发表时间：
2008-12-15
期刊：
影响因子：
4.6
通讯作者：
Kuo, Calvin J.
中科院分区：
文献类型：
--
作者：
Kuhnert, Frank;Mancuso, Michael R.;Kuo, Calvin J.
内含子microrna已被提出使小鼠基因敲除研究的设计和解释复杂化。内皮表达的Egfl7/miR-126位点在Egfl7内含子7中含有miR-126，血管生成缺陷先前被归因于Egfl7基因陷阱和lacZ敲入小鼠。令人惊讶的是，选择性修饰的Egfl7（Delta）和miR-126（Delta）等位基因显示，Egfl7（Delta/Delta）小鼠表型正常，而miR-126（Delta/Delta）小鼠携带289-nt微缺失，再现了先前描述的Egfl7胚胎和出生后视网膜血管表型。miR- 126对血管生成的调节作用通过内皮特异性缺失和成人角膜微袋实验得到证实。此外，miR-126缺失分别通过抑制PI3激酶和Spred1的p85 β亚基抑制vegf依赖性Akt和Erk信号传导。这些研究证明了内皮miRNA对血管生成的调节，将先前描述的Egfl7血管表型归因于miR- 126，并记录了无意中的miRNA失调是小鼠基因敲除策略的并发症。
Intronic microRNAs have been proposed to complicate the design and interpretation of mouse knockout studies. The endothelial-expressed Egfl7/miR-126 locus contains miR-126 within Egfl7 intron 7, and angiogenesis deficits have been previously ascribed to Egfl7 gene-trap and lacZ knock-in mice. Surprisingly, selectively floxed Egfl7(Delta) and miR-126(Delta) alleles revealed that Egfl7(Delta/Delta) mice were phenotypically normal, whereas miR-126(Delta/Delta) mice bearing a 289-nt microdeletion recapitulated previously described Egfl7 embryonic and postnatal retinal vascular phenotypes. Regulation of angiogenesis by miR- 126 was confirmed by endothelial-specific deletion and in the adult cornea micropocket assay. Furthermore, miR-126 deletion inhibited VEGF-dependent Akt and Erk signaling by derepression of the p85 beta subunit of PI3 kinase and of Spred1, respectively. These studies demonstrate the regulation of angiogenesis by an endothelial miRNA, attribute previously described Egfl7 vascular phenotypes to miR- 126, and document inadvertent miRNA dysregulation as a complication of mouse knockout strategies.