Departamento de Zoología Genética y Antropología Física, Facultad de Veterinaria, Universidade de Santiago de Compostela, Campus de Lugo, 27002 Lugo, Spain.
Genes (Basel). 2024 Sep 3;15(9):1164. doi: 10.3390/genes15091164.
Previous research has highlighted significant phenotypic discrepancies between knockout and knockdown approaches in zebrafish, raising concerns about the reliability of these methods. However, our study suggests that these differences are not as pronounced as was once believed. By carefully examining the roles of maternal and zygotic gene contributions, we demonstrate that these factors significantly influence phenotypic outcomes, often accounting for the observed discrepancies. Our findings emphasize that morpholinos, despite their potential off-target effects, can be effective tools when used with rigorous controls. We introduce the concept of graded maternal contribution, which explains how the uneven distribution of maternal mRNA and proteins during gametogenesis impacts phenotypic variability. Our research categorizes genes into three types-susceptible, immune, and "Schrödinger" (conditional)-based on their phenotypic expression and interaction with genetic compensation mechanisms. This distinction provides new insights into the paradoxical outcomes observed in genetic studies. Ultimately, our work underscores the importance of considering both maternal and zygotic contributions, alongside rigorous experimental controls, to accurately interpret gene function and the mechanisms underlying disease. This study advocates for the continued use of morpholinos in conjunction with advanced genetic tools like CRISPR/Cas9, stressing the need for a meticulous experimental design to optimize the utility of zebrafish in genetic research and therapeutic development.
先前的研究强调了在斑马鱼中，敲除和敲低方法之间存在显著的表型差异，这引起了人们对这些方法可靠性的关注。然而，我们的研究表明，这些差异并不像以前认为的那么明显。通过仔细研究母源和合子基因贡献的作用，我们证明这些因素显著影响表型结果，常常可以解释观察到的差异。我们的研究结果强调，尽管 morpholino 具有潜在的脱靶效应，但如果使用严格的对照，它们仍然是有效的工具。我们引入了分级母源贡献的概念，该概念解释了配子发生过程中母源 mRNA 和蛋白质的不均匀分布如何影响表型变异性。我们根据表型表达和与遗传补偿机制的相互作用，将基因分为敏感型、免疫型和“薛定谔”（条件型）三种类型。这种区分提供了对遗传研究中观察到的矛盾结果的新见解。最终，我们的研究强调了考虑母源和合子贡献以及严格的实验对照的重要性，以准确解释基因功能和疾病的潜在机制。本研究主张继续使用 morpholino，并结合 CRISPR/Cas9 等先进的遗传工具，强调需要精心设计实验，以优化斑马鱼在遗传研究和治疗开发中的应用。
