A gene-targeting approach for functional characterization of KIAA genes encoding extremely large proteins
A gene-targeting approach for functional characterization of KIAA genes encoding extremely large proteins
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DOI：
10.1096/fj.06-5952fje
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发表时间：
2006-08-01
期刊：
影响因子：
4.8
通讯作者：
Ohara, Osamu
中科院分区：
文献类型：
--
作者：
Nakayama, Manabu;Iida, Midori;Ohara, Osamu
鉴于哺乳动物基因组中存在数千个基因，需要标准来优先考虑其功能分析，并降低产生缺乏明显表型的基因靶向小鼠的可能性。如果集中在编码大蛋白质的基因上，初步的分析工作可能是富有成效的，因为至少有一些大蛋白质作为蛋白质复合物复杂组装的框架，它们的失活将呈现明确的、可观察的表型。在这里，我们描述了五个人类KIAA基因（KIAA1409，KIAA1440，KIAA1447，KIAA1768和KIAA1276），编码大蛋白的小鼠同源物的功能特性。基因靶向小鼠具有表型和发育缺陷，这是由于这五个基因中的三个基因的功能缺失造成的。靶向破坏KIAA1409的小鼠缺乏饮水能力，而靶向破坏KIAA1447的小鼠表现出后腿运动功能障碍。KIAA1440的破坏导致胚泡阶段的胚胎死亡。我们的方法的高成功率证明了使用反向遗传学对小鼠中的大蛋白进行全基因组功能检查的基本原理。
Given that thousands of genes exist in the mammalian genome, criteria are needed to prioritize their functional analysis and to decrease the likelihood of producing gene-targeted mice that lack overt phenotypes. Initial analysis efforts are likely to be fruitful if focused on genes encoding large proteins, since at least some large proteins serve as frameworks for intricate assembly of protein complexes, and their inactivation would render definitive, observable phenotypes. Here, we describe the functional characterization of the murine homologues of five human KIAA genes (KIAA1409, KIAA1440, KIAA1447, KIAA1768, and KIAA1276) that encode large proteins. Gene-targeted mice had phenotypic and developmental defects resulting from the functional deletion of three of these five genes. Mice with targeted disruption of KIAA1409 lacked the ability to drink, and those with targeted disruption of KIAA1447 displayed hind leg motor dysfunction. Disruption of KIAA1440 led to embryonic lethality at the blastocyst stage. The high success rate of our approach demonstrates the rationale for the genome-wide functional examination of large proteins in mice using reverse genetics.