The Serine-Threonine Protein Kinase PAK4 Is Dispensable in Zebrafish: Identification of a Morpholino-Generated Pseudophenotype
The Serine-Threonine Protein Kinase PAK4 Is Dispensable in Zebrafish: Identification of a Morpholino-Generated Pseudophenotype
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DOI：
10.1371/journal.pone.0100268
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发表时间：
2014-06-19
期刊：
影响因子：
3.7
通讯作者：
Sargent, Thomas D.
中科院分区：
文献类型：
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作者：
Law, Sheran H. W.;Sargent, Thomas D.
基于TALEN的斑马鱼pak4基因失活导致胚胎和成鱼表现出正常和可育。这与我们先前发表的基于使用反义吗啉代寡核苷酸（MO）的研究相反。我们已经排除了潜在的解释，如基因复制，选择性剪接，翻译的隐蔽启动，和非依赖于转录的RNA功能。我们的结论是pak4在斑马鱼中是不稳定的，即使得到了强有力的控制，如RNA拯救的证实，MO也可能引起误导性的假表型，这些假表型与基因突变所获得的结果不一致，因此应谨慎使用。
TALEN-based inactivation of the zebrafish pak4 gene resulted in embryos and adult fish that appear normal and fertile. This is in contrast to our previously published studies which were based on the use of antisense morpholino oligonucleotides (MOs). We have excluded potential explanations such as gene duplication, alternate splicing, cryptic initiation of translation, and translation-independent RNA function. Our conclusion is that pak4 is dispensable in zebrafish, and that even when corroborated by robust controls, such as RNA rescue, MOs may elicit misleading pseudophenotypes that do not correspond to results obtained by genetic mutations, and should thus be used with caution.